Oxindoles as kinase inhibitors

ABSTRACT

The present invention relates to oxindoles of the formula I, their use as protein kinase activators or inhibitors, a method for their manufacture, their use for the preparation of a medicament for the treatment of diseases and their use for the manufacture of a pharmaceutical composition.

FIELD OF THE INVENTION

The present invention relates to oxindoles, their use as protein kinaseactivators or inhibitors, a method for their manufacture, their use forthe preparation of a medicament for the treatment of diseases and theiruse for the manufacture of a pharmaceutical composition.

BACKGROUND OF THE INVENTION

Protein kinases are involved in signaling pathways for such importantcellular activities as responses to extracellular signals and cell cyclecheckpoints. Inhibition or activation of specific protein kinasesprovides a means of intervening in these signaling pathways, for exampleto block the effect of an extracellular signal, to release a cell fromcell cycle checkpoint, etc. Defects in the activity of protein kinasesare associated with a variety of pathological or clinical conditions,where there is a defect in the signaling mediated by protein kinases.Such conditions include those associated with defects in cell cycleregulation or in response to extracellular signals, e.g., immunologicaldisorders, autoimmune and immunodeficiency diseases; hyperproliferativedisorders, which may include psoriasis, arthritis, inflammation,endometriosis, scarring, cancer, etc.

Aberrant activation of the Ras-Raf-MAP kinase signaling pathwaycontributes to the process of tumorigenesis, i.e. the conversion of anormal into a malignant cell. Many molecular details have been workedout, how the MAPK signaling module initiates proliferation or inhibitsapoptotic response, thus explaining how the homeostatic balance betweenproliferation and cell death can be disturbed. MKK1 and MKK2, also knownas MEK1 and MEK2, represent a key element in this signaling cascade.MEKs are activated by Raf kinase phosphorylation and, in turn,phosphorylate and activate their bona fide substrates ERK1 and ERK2.Oncogenic Ras mutations, mutations in the effector kinase B-Raf and evengrowth factor overexpression and mutation may lead to persistentactivation of the MAPK pathway justifying the potential of MAPK pathwayinhibitors at a level of Rat, MEK or ERK as promising anticancertherapeutics.

Therefore, compounds, which are active in modulating purified kinaseproteins, e.g. there is a modulation in the phosphorylation of aspecific substrate in the presence of the compound, can be used for thetreatment of protein kinase-dependent diseases and conditions, such ascancer, tumour growth, artherosclerosis, age-related maculardegeneration, diabetic retinopathy, inflammatory diseases and the like,in mammals.

Amino-oxindoles are known from e.g. WO9952869, WO9962882, WO04026829,WO04009546, WO04009547, EP104860, U.S. Pat. No. 4,145,422, WO03027102and WO0149287.3-(1-Amino-2-phenyl-ethylidene)-1-methyl-1,3-dihydro-indol-2-one isknown from Wenkert, E. et al., J. Am. Chem. Soc. (1958), 80, 4899-4903,3-(Amino-phenyl-methylene)-1,3-dihydro-indol-2-one is known from Stauss,U. et al., Helv. Chim. Acta (1972), 55(3), 771-780.

Compounds described as putative MEK inhibitors are known from e.g.WO03062191, WO04056789, WO03077914, WO03077855, WO04041811 andWO04048386.

Thus, as there remains a need in advantageous therapeutics, a preferredobject of the present invention was to provide new pharmaceuticallyactive compounds. A particularly preferred aim of the present inventionwas to provide effective modulators of one or more protein kinases,especially selected from the group of Raf, MEK, PKB, Tie2, PDGFR, Met,SGK1, IGF1R and VEGFR.

BRIEF DESCRIPTION OF THE INVENTION

Surprisingly, the compounds of the general formula I show pharmaceuticalactivities as they act as effective modulators (activators orinhibitors) of one or more protein kinases selected from the group ofRaf, MEK, PKB, Tie2, PDGFR, Met, SGK1, IGF1R and VEGFR.

Therefore, an embodiment of the present invention are compounds of theformula I,

wherein

-   X is (CH₂)_(p),-   R¹ is Ar or Het,-   R² is H, A, Ar, (CH₂)_(m)CON(R⁸)₂, (CH₂)_(m)CONHAr, S(O)_(m)A,    S(O)_(m)Ar, NHCOA, NHCOAr, NHSO₂A, NHSO₂Ar, SO₂N(R⁸)₂,    N(CH₂)_(n)C(CH₃)₂(CH₂)_(n)N(R⁸)₂, (CH₂)_(n)N(R⁸)SO_(m)A,    (CH₂)_(n)N(R⁸)SO_(m)Ar, (CH₂)_(n)SO_(m)A, (CH₂)_(n)SO_(m)Ar or    (CH₂)_(n)SO_(m)N(R⁸)A,-   R³, R⁴, R⁵, R⁶, R⁷ are independently from each other H, A, Ar,    OR^(B), SR⁸, OAr, SAr, N(R⁸)₂, NHAr, NAr₂, Hal, NO₂, CN, COW, COAr,    S(O)_(m)A, S(O)_(m)Ar, NHCOA, NHCOAr, NHSO₂A, NHSO₂Ar, SO₂N(R⁸)₂,    O(CH₂)_(n)N(R⁸)₂, O(CH₂)_(n)NHR⁸, O(CH₂)_(n)-morpholine,    O(CH₂)_(n)-piperazine, O(CH₂)_(n)-pyrrolidine,    O(CH₂)_(n)-piperidine, O-piperidine, O(CH₂)_(n)-oxopiperazine,    O(CH₂)_(n)-oxomorpholine, O(CH₂)_(n)-oxopyrrolidine,    O(CH₂)_(n)C(CH₃)₂(CH₂)_(n)N(R⁸)₂, N(CH₂)_(n)C(CH₃)₂(CH₂)_(n)N(R⁸)₂,    O(CH₂)_(n)N(R⁸)SO_(m)A, O(CH₂)_(n)N(R⁸)SO_(m)Ar,    O(CH₂)_(n)N(R⁸)SO_(m)N(R⁸)₂A, (CH₂)_(n)N(R⁸)SO_(m)A,    (CH₂)_(n)N(R⁸)SO_(m)Ar, (CH₂)_(n)N(R⁸)SO_(m)N(R⁸)₂A,    O(CH₂)_(n)SO_(m)A, O(CH₂)_(n)SO_(m)Ar, O(CH₂)_(n)SO_(m)N(R⁸)A,    (CH₂)_(n)SO_(m)A, (CH₂)_(n)SO_(m)Ar or (CH₂)_(n)SO_(m)N(R⁸)A,-   R⁸ is H, A or A-Ar,-   A is a linear or branched alkyl or a cycloalkyl which is optionally    substituted by Hal,-   Ar is aryl,-   Het is heteroaryl,-   Hal Cl, Br, I or F,-   n, p are independently from each other 0-5,-   m is 0-2,    with the provisio, that one of the residues R², R³, R⁴, R⁵, R⁶ or R⁷    is other than H and that    3-(1-Amino-2-phenyl-ethylidene)-1-methyl-1,3-dihydro-indol-2-one is    excluded, and the physiologically acceptable salts, derivatives,    prodrugs, solvates and stereoisomers thereof, including mixtures    thereof in all ratios.

A further preferred embodiment of the present invention are compoundsaccording to formula I, wherein

-   X is (CH₂)_(p),-   R¹ is Ar or Het,-   R² is H, A, Ar, (CH₂)_(m)CON(R⁸)₂, (CH₂)_(m)CONHAr,    (CH₂)_(n)N(R⁸)SO_(m)A, (CH₂)_(n)N(R⁸)SO_(m)Ar or    (CH₂)_(n)SO_(m)N(R⁸)A,-   R³, R⁴, R⁵, R⁶, R⁷ are independently from each other H, A, Ar, OR⁸,    SR⁸, OAr, SAr, N(R⁸)₂, NHAr, NAr₂, Hal, NO₂, ON, COR^(S), COAr,    NHCOA, NHCOAr, NHSO₂A, NHSO₂Ar, SO₂N(R⁸)₂, O(CH₂)_(n)N(R⁸)₂,    O(CH₂)_(n)NHR⁸, O(CH₂)_(n)-morpholine, O(CH₂)_(n)-piperazine,    O(CH₂)_(n)-pyrrolidine, O(CH₂)_(n)-piperidine, O-piperidine,    O(CH₂)_(n)-oxopiperazine, O(CH₂)_(n)-oxomorpholine,    O(CH₂)_(n)-oxopyrrolidine, O(CH₂)_(n)C(CH₃)₂(CH₂)_(n)N(R⁸)₂ or    N(CH₂)_(n)C(CH₃)₂(CH₂)_(n)N(R⁸)₂,-   R⁸ is H, A or A-Ar,-   A is a linear or branched alkyl or a cycloalkyl which is optionally    substituted by Hal,-   Ar is aryl,-   Het is heteroaryl,-   Hal Cl, Br, I or F,-   n, p are independently from each other 0-5,-   m is 0-2,    with the provisio, that one of the residues R², R³, R⁴, R⁵, R⁶ or R⁷    is other than H and that    3-(1-Amino-2-phenyl-ethylidene)-1-methyl-1,3-dihydro-indol-2-one is    excluded, and the physiologically acceptable salts, derivatives,    prodrugs, solvates and stereoisomers thereof, including mixtures    thereof in all ratios.

A further preferred embodiment of the present invention are compoundsaccording to formula I, wherein

-   X is (CH₂)_(p),-   R¹ is Ar or Het,-   R² is H, A, Ar, (CH₂)_(m)CON(R⁸)₂ or (CH₂)_(m)CONHAr,-   R³, R⁴, R⁵, R⁶, R⁷ are independently from each other H, A, Ar, OR⁸,    SR⁸, OAr, SAr, N(R⁸)₂, NHAr, NAr₂, Hal, NO₂, CN, COR⁸, COAr, NHCOA,    NHCOAr, NHSO₂A, NHSO₂Ar, SO₂N(R⁸)₂, O(CH₂)_(n)N(R⁸)₂ or    O(CH₂)_(n)NHR⁸,-   R⁸ is H, A or A-Ar,-   A is a linear or branched alkyl or a cycloalkyl which is optionally    substituted by Hal,-   Ar is aryl,-   Het is heteroaryl,-   Hal Cl, Br, I or F,-   n, p are independently from each other 0-5,-   m is 0-2,    with the provisio, that one of the residues R², R³, R⁴, R⁵, R⁶ or R⁷    is other than H and that    3-(1-Amino-2-phenyl-ethylidene)-1-methyl-1,3-dihydro-indol-2-one is    excluded, and the physiologically acceptable salts, derivatives,    prodrugs, solvates and stereoisomers thereof, including mixtures    thereof in all ratios.

A further preferred embodiment of the present invention are compoundsaccording to formula I, wherein

-   X is (CH₂)_(p),-   p is 0-5,-   R¹ is Ar or Het,-   R², R³, R⁴, R⁶, R⁷ are H,-   R⁵ is Hal,    and the physiologically acceptable salts, derivatives, prodrugs,    solvates and stereoisomers thereof, including mixtures thereof in    all ratios.

A further preferred embodiment of the present invention are compoundsaccording to formula I, wherein

-   X is (CH₂)_(p),-   p is 0-5,-   R¹ is Ar or Het,-   R², R³, R⁴, R⁶, R⁷ are H,-   R⁴ is Hal,    and the physiologically acceptable salts, derivatives, prodrugs,    solvates and stereoisomers thereof, including mixtures thereof in    all ratios.

A further preferred embodiment of the present invention are compoundsaccording to formula I, wherein

-   X is (CH₂)_(p),-   p is 0-5,-   R¹ is Ar or Het,-   R², R³, R⁴, R⁶, R⁷ are H,-   R², is A or Ar,    with the provisio, that    3-(1-Amino-2-phenyl-ethylidene)-1-methyl-1,3-dihydroindol-2-one is    excluded, and the physiologically acceptable salts, derivatives,    prodrugs, solvates and stereoisomers thereof, including mixtures    thereof in all ratios.

A further preferred embodiment of the present invention are compoundsaccording to formula I, wherein

-   X is (CH₂)_(p),-   p is 0-5,-   R¹ is Ar or Het,-   R², R³, R⁴, R⁶ are H,-   R⁵ is Hal,-   R⁷ is Hal or OR⁸,-   R⁸ is H or A,    and the physiologically acceptable salts, derivatives, prodrugs,    solvates and stereoisomers thereof, including mixtures thereof in    all ratios.

A further preferred embodiment of the present invention are compoundsaccording to formula I, wherein

-   X is (CH₂)_(p),-   p is 0-5,-   R¹ is Ar or Het,-   R², R³, R⁵, R⁶ are H,-   R⁴ is Hal,-   R⁷ is Hal or OR⁸,-   R⁸ is H or A,    and the physiologically acceptable salts, derivatives, prodrugs,    solvates and stereoisomers thereof, including mixtures thereof in    all ratios.

A further preferred embodiment of the present invention are compoundsaccording to formula I, wherein

-   X is (CH₂)_(p),-   p is 0-5,-   R¹ is Ar or Het,-   R², R³, R⁴, R⁶ are H,-   R⁵ is Hal,-   R⁷ is Hal or OR⁸,-   R⁸ is H or A,    and the physiologically acceptable salts, derivatives, prodrugs,    solvates and stereoisomers thereof, including mixtures thereof in    all ratios.

A further preferred embodiment of the present invention are compoundsaccording to formula I, wherein

-   X is (CH₂)_(p),-   p is 0,-   R¹ is phenyl,-   R², R³, R⁴, R⁶ are H,-   R² is A or Ar,-   R⁷ is Hal or OR⁸,-   R⁸ is H or A,    and the physiologically acceptable salts, derivatives, prodrugs,    solvates and stereoisomers thereof, including mixtures thereof in    all ratios.

An especially preferred embodiment of the present invention arecompounds according to formula I, selected from the group consisting of

-   a) 3-(Amino-phenyl-methylene)-6-chloro-1,3-dihydro-indol-2-one,-   b) 3-(Amino-phenyl-methylene)-5-chloro-1,3-dihydro-indol-2-one,-   c) 3-[Amino-(4-hydroxy-phenyl)-methylene]-1,3-dihydro-indol-2-one-   d) 3-[Amino-(4-iodo-phenyl)-methylene]-1,3-dihydro-indol-2-one-   e)    3-(Amino-(4-iodo-phenyl)-methylene)-6-chloro-1,3-dihydro-indol-2-one,-   f)    3-(Amino-(4-iodo-phenyl)-methylene)-5-chloro-1,3-dihydro-indol-2-one,-   g) 3-[Amino-(3-iodo-phenyl)-methylene]-1,3-dihydro-indol-2-one-   h)    3-(Amino-(3-iodo-phenyl)-methylene)-6-chloro-1,3-dihydro-indol-2-one,-   i)    3-(Amino-(3-iodo-phenyl)-methylene)-5-chloro-1,3-dihydro-indol-2-one,-   j)    3-(Amino-(3-iodo-phenyl)-methylene)-1-methyl-1,3-dihydro-indol-2-one,-   k) 3-(Amino-phenyl-methylene)-5-bromo-1,3-dihydro-indol-2-one,

l) 3-(Amino-(4-iodo-phenyl)-methylene)-1-methyl-1,3-dihydro-indol-2-one,

-   m)    3-(Amino-(4-iodo-phenyl)-methylene)-5-bromo-1,3-dihydro-indol-2-one-   n)    3-(Amino-(3-iodo-phenyl)-methylene)-5-bromo-1,3-dihydro-indol-2-one,-   o) 3-[Amino-(4-methoxy-phenyl)methylene]-1,3-dihydro-indol-2-one-   p)    3-(Amino-(4-methoxy-phenyl)-methylene)-6-chloro-1,3-dihydro-indol-2-one,-   q)    3-(Amino-(4-methoxy-phenyl)-methylene)-5-bromo-1,3-dihydro-indol-2-one,-   r)    3-(Amino-(4-hydroxy-phenyl)-methylene)-6-chloro-1,3-dihydro-indol-2-one,-   s)    3-(Amino-(4-hydroxy-phenyl)-methylene)-5-chloro-1,3-dihydro-indol-2-one,-   t) 3-(Amino-phenyl-methylene)-1-methyl-1,3-dihydro-indol-2-one,-   u)    3-(Amino-(4-hydroxy-phenyl)-methylene)-1-methyl-1,3-dihydro-indol-2-one,-   v)    3-(Amino-(4-hydroxy-phenyl)-methylene)-5-bromo-1,3-dihydro-indol-2-one,-   w)    3-(Amino-(4-methoxy-phenyl)-methylene)-5-chloro-1,3-dihydro-indol-2-one,-   x) 3-[Amino-(4-fluoro-phenyl)-methylene]-1,3-dihydro-indol-2-one-   y)    3-(Amino-(4-fluoro-phenyl)-methylene)-5-chloro-1,3-dihydro-indol-2-one,-   z)    3-(Amino-(4-methoxy-phenyl)-methylene)-1-methyl-1,3-dihydro-indol-2-one,    and the physiologically acceptable salts, derivatives, prodrugs,    solvates and stereoisomers thereof, including mixtures thereof in    all ratios.

Suitable salts and pharmaceutically acceptable salts of the compoundsaccording to the invention are conventional non-toxic salts and includeacid addition salts such as organic acid salts (e.g. acetate,trifluoroacetate, maleate, tartrate, methanesulfonate, benzenesulfonate,formate, toluenesulfonate), inorganic acid salt (e.g. hydrochloride,hydrobromide, hydroiodide, sulfate, nitrate, phosphate), or salts withan amino acid (e.g. arginine, aspartic acid, glutamic acid), or metalsalts such as alkali metal salts (e.g. sodium salt, potassium salt) andalkaline earth metal salts (e.g. calcium salt, magnesium salt), ammoniumsalts, or organic base salts (e.g. trimethylamine salt, triethylaminesalt, pyridine salt, picoline salt, dicyclohexylamine salt,N,N′-dibenzylethylenediamine salt).

The term “pharmaceutically usable derivatives” or “pharmaceuticallyacceptable derivatives” is taken to mean, for example, the salts of thecompounds according to the invention and so-called prodrug compounds.The term refers to any pharmaceutically acceptable derivative of acompound of the present invention, for example, an ester or an amide,which upon administration to a mammal is capable of providing (directlyor indirectly) a compound of the present invention or an activemetabolite thereof. Such derivatives are clear to those skilled in theart, without undue experimentation, and with reference to the teachingof Burger's Medicinal Chemistry And Drug Discovery, 5th Edition, Vol 1:Principles and Practice, which is incorporated herein by reference tothe extent that it teaches physiologically functional derivatives.

The term “prodrug derivatives” is taken to mean, for example, compoundsof the present invention which have been modified, for example, withalkyl or acyl groups, sugars or oligopeptides and which are rapidlycleaved in the organism and thus release the active ingredientsaccording to the invention. These also include biodegradable polymerderivatives of the compounds according to the invention, as described,for example, in Int. J. Pharm. 115, 61-67 (1995).

The invention also relates to mixtures of the compounds according to theinvention, for example mixtures of two diastereomers, for example in theratio 1:1, 1.2, 1:3, 1:4, 1:5, 1:10, 1:100 or 1:1000. These areparticularly preferably mixtures of stereoisomeric compounds.

Additionally, the invention comprises the polymorphic forms of thecompounds according to the invention, e.g. the amorphic and crystallinepolymorphic forms.

As used herein, the term “solvate” preferably refers to a complex ofvariable stoichiometry formed by a solute and a solvent. The termsolvates of the compounds is therefore taken to mean adductions of inertsolvent molecules onto the compounds, which form owing to their mutualattractive force. Such solvents for the purpose of the invention may notinterfere with the biological activity of the solute. Preferably thesolvent used is a pharmaceutically acceptable solvent. Examples ofsuitable pharmaceutically acceptable solvents include, withoutlimitation, water, ethanol and acetic acid. Most preferably the solventused is water. Solvates are, for example, monohydrates, dihydrates oralcoholates.

Certain of the compounds described herein may contain one or more chiralatoms, or may otherwise be capable of existing as two or morestereoisomers, which are usually enantiomers and/or diastereomers.Accordingly, the compounds of this invention include mixtures ofstereoisomers, especially mixtures of enantiomers, as well as purifiedstereoisomers, especially purified enantiomers, or stereoisomericallyenriched mixtures, especially enantiomerically enriched mixtures. Alsoincluded within the scope of the invention are the individual isomers ofthe compounds of the present invention as well as any wholly orpartially equilibrated mixtures thereof. The present invention alsocovers the individual isomers of the compounds represented by theformulas above as mixtures with isomers thereof in which one or morechiral centers are inverted. Also, it is understood that all tautomersand mixtures of tautomers of the compounds of the present invention areincluded within the scope of the compounds of the present invention andpreferably the formulae and subformulae corresponding thereto.

Racemates obtained can be resolved into the isomers mechanically orchemically by methods known per se. Diastereomers are preferably formedfrom the racemic mixture by reaction with an optically active resolvingagent. Examples of suitable resolving agents are optically active acids,such as the D and L forms of tartaric acid, diacetyltartaric acid,dibenzoyltartaric acid, mandelic acid, malic acid, lactic acid or thevarious optically active camphorsulfonic acids, such asp-camphorsulfonicacid. Also advantageous is enantiomer resolution with the aid of acolumn filled with an optically active resolving agent (for exampledinitrobenzoylphenylglycine); an example of a suitable eluent is ahexane/isopropanol/acetonitrile mixture.

The diastereomer resolution can also be carried out by standardpurification processes, such as, for example, chromatography orfractional crystallization.

It is of course also possible to obtain optically active compounds ofthe present invention by the methods described above by using startingmaterials which are already optically active.

Unless indicated otherwise, it is to be understood that reference to thecompounds of the present invention preferably includes the reference tothe subformulae corresponding thereto. It is also understood that thefollowing embodiments, including uses and compositions, although recitedwith respect to the compounds of the present invention are preferablyalso applicable to subformulae.

As used herein, the terms “group”, “residue” and “radical” or “groups”,“residues” and “radicals” are usually used as synonyms, respectively, asit is common practice in the art.

As used herein, the term “optionally” means that the subsequentlydescribed event(s) may or may not occur, and includes both event(s),which occur, and events that do not occur.

As used herein, the term “substituted” preferably refers to substitutionwith the named substituent or substituents, multiple degrees ofsubstitution being allowed unless otherwise stated.

Subject of the present invention are especially compounds of the presentinvention in which one or more substituents or groups, preferably themajor part of the substituents or groups has a meaning which isindicated as preferred, more preferred, even more preferred orespecially preferred.

As used herein, the term “halogen” or “hal” preferably refers tofluorine (F), chlorine (Cl), bromine (Br) or iodine (I).

As used herein, the term “A” or “alkyl” preferably refers to a straightor branched chain hydrocarbon having from one to twelve carbon atoms,wherein optionally 1-5H atoms are replaced by F and/or Cl, multipledegrees of substitution being allowed. Examples of “alkyl” as usedherein include, but are not limited to, methyl, ethyl, n-propyl,isopropyl, n-butyl, isobutyl, t-butyl, n-pentyl, isopentyl, and thelike.

As used herein, the term “cycloalkyl” preferably refers to anon-aromatic cyclic hydrocarbon ring system, with one or more ringsattached to each other, each ring preferably having from three to sevencarbon atoms, which optionally includes an alkyl linker, preferably aC₁-C₆ alkyl linker, through which it may be attached. Optionally, in the“cycloalkyl” 1-5H atoms are replaced by F and/or Cl, multiple degrees ofsubstitution being allowed. The alkyl or C₁-C₆ alkyl group is as definedabove. Exemplary “cycloalkyl” groups include, but are not limited to,cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and cycloheptyl.

As used herein, the term “Ar” or “aryl” preferably refers to a benzenering or a benzene ring system, which is optionally substituted by A, Ar,OR⁸, SR⁸, OAr, SAr, N(R⁸)₂, NHAr, NAr₂, Hal, NO₂, ON, COR⁸, COAr,S(O)_(m)A, S(O)_(m)Ar, NHCOA, NHCOAr, NHSO₂A, NHSO₂Ar, SO₂N(R⁸)₂,O(CH₂)_(n)N(R⁸)₂, O(CH₂)_(n)NHR⁸, O(CH₂)_(n)-morpholine,O(CH₂)_(n)-piperazine, O(CH₂)_(n)-pyrrolidine, O(CH₂)_(n)-piperidine,O-piperidine, O(CH₂)_(n)-oxopiperazine, O(CH₂)_(n)-oxomorpholine,O(CH₂)_(n)-oxopyrrolidine, O(CH₂)_(n)C(CH₃)₂(CH₂)_(n)N(R⁸)₂,N(CH₂)_(n)C(CH₃)₂(CH₂)_(n)N(R⁸)₂, O(CH₂)_(n)N(R⁸)SO_(m)A,O(CH₂)_(n)N(R⁸)SO_(m)Ar, O(CH₂)_(n)N(R⁸)SO_(m)N(R⁸)₂A,(CH₂)_(n)N(R⁸)SO_(m)A, (CH₂)_(n)N(R⁸)SO_(m)Ar,(CH₂)_(n)N(R⁸)SO_(m)N(R⁸)₂A, O(CH₂)_(n)SO_(m)A, O(CH₂)_(n)SO_(m)Ar,O(CH₂)_(n)SO_(m)N(R⁸)A, (CH₂)_(n)SO_(m)A, (CH₂)_(n)SO_(m)Ar,(CH₂)_(n)SO_(m)N(R⁸)A, and the like, multiple degrees of substitutionbeing allowed. Examples of “aryl” groups include, but are not limited toPhenyl, 2-naphthyl, 1-naphthyl, biphenyl, anthracyl, phenanthracyl, aswell as substituted derivatives thereof.

As used herein, the term “Het” or the term “heteroaryl” preferablyrefers to a monocyclic five to seven-membered aromatic ring, or to afused bicyclic aromatic ring system comprising two of such monocyclicfive to seven-membered aromatic rings. These heteroaryl rings containone or more nitrogen, sulfur and/or oxygen heteroatoms, where N-oxidesand sulfur oxides and dioxides are permissible heteroatom substitutionsand may be optionally substituted by one or more substituents, selectedfrom the group consisting of A, Ar, OR⁸, SR⁸, OAr, SAr, N(R⁸)₂, NHAr,NAr₂, Hal, NO₂, ON, COR⁸, COAr, S(O)_(m)A, S(O)_(m)Ar, NHCOA, NHCOAr,NHSO₂A, NHSO₂Ar, SO₂N(R⁸)₂, O(CH₂)_(n)N(R⁸)₂, O(CH₂)_(n)NHR⁸,O(CH₂)_(n)-morpholine, O(CH₂)_(n)-piperazine, O(CH₂)_(n)-pyrrolidine,O(CH₂)_(n)-piperidine, O-piperidine, O(CH₂)_(n)-oxopiperazine,O(CH₂)_(n)-oxomorpholine, O(CH₂)_(n)-oxopyrrolidine,O(CH₂)_(n)C(CH₃)₂(CH₂)_(n)N(R⁸)₂, N(CH₂)_(n)C(CH₃)₂(CH₂)_(n)N(R⁸)₂,O(CH₂)_(n)N(R⁸)SO_(m)A, O(CH₂)_(n)N(R⁸)SO_(m)Ar,O(CH₂)_(n)N(R⁸)SO_(m)N(R⁸)₂A, (CH₂)_(n)N(R⁸)SO_(m)A,(CH₂)_(n)N(R⁸)SO_(m)Ar, (CH₂)_(n)N(R⁸)SO_(m)N(R⁸)₂A, O(CH₂)_(n)SO_(m)A,O(CH₂)_(n)SO_(m)Ar, O(CH₂)_(n)SO_(m)N(R⁸)A, (CH₂)_(n)SO_(m)A,(CH₂)_(n)SO_(m)Ar, (CH₂)_(n)SO_(m)N(R⁹)A, and the like. Examples of“heteroaryl” moieties include, but are not limited to furanyl,thiophenyl, pyranyl, pyrrolyl, imidazolyl, pyrazolyl, thiazolyl,pyridinyl, pyrazinyl, quinolinyl, isoquinolinyl, triazolyl, tetrazolyl,oxazolyl, isoxazolyl, oxadiazolyl, oxo-pyridyl, thiadiazolyl,isothiazolyl, pyridazyl, pyrimidinyl, benzofuranyl, benzothiophenyl,indolyl, indazolyl, and the like.

The nomenclature as used herein for defining compounds, especially thecompounds according to the invention, is in general based on the rulesof the IUPAC-organisation for chemical compounds and especially organiccompounds.

A further preferred embodiment of the present invention is a method forthe manufacture of a compound according to formula I, characterized inthat

-   a) an aromatic or heteroaromatic nitrile according to formula II,

-   -   wherein R¹, Wand X are as defined above, is reacted with a        straight or branched chain alcohol according to formula III,

HO—R⁹  III

-   -   wherein R⁹ is (CH₂)_(q) and q is 1-10,    -   and that the product according to formula IV,

-   -   wherein R¹, R⁷, R⁹ and X are as defined above, is then reacted        with a compound according to formula V,

-   -   wherein R², R³, R⁴, R⁵ and R⁶ are as defined above, or

-   b) a compound of formula I is isolated and/or treated with an acid    or a base, to obtain the salt thereof.

A physiologically acceptable salt of a compound according to formula Ican also be obtained by isolating and/or treating the compound offormula I obtained by the described reaction with an acid or a base.

For a further detailed description of the manufacturing processes,please see also example 1 and the following general description of thepreferred conditions.

All crude products were subjected to standard chromatography usingsolvent mixtures containing methanol, ethanol, isopropanol, n-hexane,cyclohexane, ethylacetat, dichlormethane or petrol ether, respectively.

The compounds of the formula I and also the starting materials for theirpreparation are prepared by methods as described in the examples or bymethods known per se, as described in the literature (for example instandard works, such as Houben-Weyl, Methoden der Organischen Chemie[Methods of Organic Chemistry], Georg Thieme Verlag, Stuttgart; OrganicReactions, John Wiley & Sons, Inc., New York), to be precise underreaction conditions which are known and suitable for the said reactions.Use can also be made here of variants which are known per se, but arenot mentioned here in greater detail.

The starting materials for the claimed process may, if desired, also beformed in situ by not isolating them from the reaction mixture, butinstead immediately converting them further into the compounds of theformula I. On the other hand, it is possible to carry out the reactionstepwise.

Preferably, the reaction of the compounds is carried out in the presenceof a suitable solvent, that is preferably inert under the respectivereaction conditions. Examples of suitable solvents are hydrocarbons,such as hexane, petroleum ether, benzene, toluene or xylene; chlorinatedhydrocarbons, such as trichlorethylene, 1,2-dichloroethane,tetrachloromethane, chloroform or dichloromethane; alcohols, such asmethanol, ethanol, isopropanol, n-propanol, n-butanol or tert-butanol;ethers, such as diethyl ether, diisopropyl ether, tetrahydrofuran (THE)or dioxane; glycol ethers, such as ethylene glycol monomethyl ormonoethyl ether or ethylene glycol dimethyl ether (diglyme); ketones,such as acetone or butanone; amines, such as triethylamine; amides, suchas acetamide, dimethylacetamide, dimethylformamide (DMF) orN-methylpyrrolidinone (NMP); nitriles, such as acetonitrile; sulfoxides,such as dimethyl sulfoxide (DMSO); nitro compounds, such as nitromethaneor nitrobenzene; esters, such as ethyl acetate, or mixtures of the saidsolvents or mixtures with water. Polar solvents are in generalpreferred. Examples for suitable polar solvents are chlorinatedhydrocarbons, alcohols, glycol ethers, nitriles, amides and sulfoxidesor mixtures thereof. More preferred are 1,4-dioxane, 1-butanol andtriethylamine.

As stated above, the reaction temperature is between about −100° C. and300° C., more preferred between 0 and 250° C., preferably including theirradiation in a microwave, depending on the reaction step and theconditions used.

Reaction times are generally in the range between some minutes andseveral days, depending on the reactivity of the respective compoundsand the respective reaction conditions. Suitable reaction times arereadily determinable by methods known in the art, for example reactionmonitoring. Based on the reaction temperatures given above, suitablereaction times generally lie in the range between 10 min and 48 hrs.

A base of the formula I can be converted into the associatedacid-addition salt using an acid, for example by reaction of equivalentamounts of the base and the acid in a preferably inert solvent, such asethanol, followed by evaporation. Suitable acids for this reaction are,in particular, those which give physiologically acceptable salts. Thus,it is possible to use inorganic acids, for example sulfuric acid,sulfurous acid, dithionic acid, nitric acid, hydrohalic acids, such ashydrochloric acid or hydrobromic acid, phosphoric acids, such as, forexample, orthophosphoric acid, sulfamic acid, furthermore organic acids,in particular aliphatic, alicyclic, araliphatic, aromatic orheterocyclic monobasic or polybasic carboxylic, sulfonic or sulfuricacids, for example formic acid, acetic acid, propionic acid, hexanoicacid, octanoic acid, decanoic acid, hexadecanoic acid, octadecanoicacid, pivalic acid, diethylacetic acid, malonic acid, succinic acid,pimelic acid, fumaric acid, maleic acid, lactic acid, tartaric acid,malic acid, citric acid, gluconic acid, ascorbic acid, nicotinic acid,isonicotinic acid, methane- or ethanesulfonic acid, ethanedisulfonicacid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid,trimethoxybenzoic acid, adamantanecarboxylic acid, p-toluenesulfonicacid, glycolic acid, embonic acid, chlorophenoxyacetic acid, asparticacid, glutamic acid, proline, glyoxylic acid, palmitic acid,parachlorophenoxyisobutyric acid, cyclohexanecarboxylic acid, glucose1-phosphate, naphthalenemono- and -disulfonic acids or laurylsulfuricacid.

Salts with physiologically unacceptable acids, for example picrates, canbe used to isolate and/or purify the compounds of the formula I.

On the other hand, compounds of the formula I can be converted into thecorresponding metal salts, in particular alkali metal salts or alkalineearth metal salts, or into the corresponding ammonium salts, using bases(for example sodium hydroxide, potassium hydroxide, sodium carbonate orpotassium carbonate). Suitable salts are furthermore substitutedammonium salts, for example the dimethyl-, diethyl- anddiisopropylammonium salts, monoethanol-, diethanol- anddiisopropanolammonium salts, cyclohexyl- and dicyclohexylammonium salts,dibenzylethylenediammonium salts, furthermore, for example, salts witharginine or lysine.

If desired, the free bases of the formula I can be liberated from theirsalts by treatment with strong bases, such as sodium hydroxide,potassium hydroxide, sodium carbonate or potassium carbonate, so long asno further acidic groups are present in the molecule. In the cases wherethe compounds of the formula I have free acid groups, salt formation canlikewise be achieved by treatment with bases. Suitable bases are alkalimetal hydroxides, alkaline earth metal hydroxides or organic bases inthe form of primary, secondary or tertiary amines.

Every reaction step described herein can optionally be followed by oneor more working up procedures and/or isolating procedures. Suitable suchprocedures are known in the art, for example from standard works, suchas Houben-Weyl, Methadon der organischen Chemie [Methods of OrganicChemistry], Georg-Thieme-Verlag, Stuttgart). Examples for suchprocedures include, but are not limited to evaporating a solvent,distilling, crystallization, fractionised crystallization, extractionprocedures, washing procedures, digesting procedures, filtrationprocedures, chromatography, chromatography by HPLC and dryingprocedures, especially drying procedures in vacuum and/or elevatedtemperature.

Proliferative diseases are caused by a defect in the intracellularsignaling system, or the signal transduction mechanism of certainproteins. One of the principal mechanisms by which cellular regulationis effected is through the transduction of extracellular signals acrossthe membrane, that in turn modulate biochemical pathways within thecell. Protein phosphorylation represents one course by whichintracellular signals are propagated from molecule to molecule resultingfinally in a cellular response. These signal transduction cascades arehighly regulated and often overlapping, as evident from the existence ofmany protein kinases as well as phosphatases. Phosphorylation ofproteins occurs predominantly at serine, threonine or tyrosine residues,and protein kinases have therefore been classified by their specificityof phosphorylation site, i.e. serine/threonine kinases and tyrosinekinases. Since phosphorylation is such a ubiquitous process within cellsand since cellular phenotypes are largely influenced by the activity ofthese pathways, it is currently believed that a number of disease statesand/or diseases are attributable to either aberrant activation orfunctional mutations in the molecular components of kinase cascades.Defects include a change either in the intrinsic activity or in thecellular concentration of one or more signaling proteins in thesignaling cascade. The cell may produce a growth factor that binds toits own receptors, resulting in an autocrine loop, which continuallystimulates proliferation. Mutations or overexpression of intracellularsignaling proteins can lead to spurious mitogenic signals within thecell. Consequently, considerable attention has been devoted to thecharacterization of kinase proteins and compounds that are able tomodulate their activity (for a review see: Weinstein-Oppenheimer et al.Pharma. &. Therap., 2000, 88: 229-279).

Tyrosine kinases are a class of enzymes, which catalyse the transfer ofthe terminal phosphate of adenosine triphosphate to tyrosine residues inprotein substrates. It is thought that tyrosine kinases, throughsubstrate phosphorylation, play a crucial role in signal transductionfor a number of cell functions. Although the precise mechanisms ofsignal transduction are still unclear, tyrosine kinases have been shownto be important contributing factors in cell proliferation,carcinogenesis and cell differentiation. Tyrosine kinases can becategorised as receptor-type tyrosine kinases or non-receptor typetyrosine kinases. Receptor-type tyrosine kinases have an extracellularportion, a transmembrane portion and an intracellular portion, whilenon-receptor type tyrosine kinases are exclusively intracellular.

Tyrosine kinases consist of a multiplicity of transmembrane receptorswith different biological activity. Thus, about 20 different subfamiliesof receptor-type tyrosine kinases have been identified. One tyrosinekinase subfamily, known as the HER subfamily, consists of EGFR, HER2,HERS and HERO. Ligands from this subfamily of receptors includeepithelial growth factor, TGF-α, amphiregulin, HB-EGF, betacellulin andheregulin. Another subfamily of these receptor-type tyrosine kinases isthe insulin subfamily, which includes INS-R, IGF-IR and IR-R. The PDGFsubfamily includes the PDGF-α and -β receptors, CSFIR, c-kit and FLK-II.In addition, there is the FLK family, which consists of the kinaseinsert domain receptor (KDR), foetal liver kinase-1 (FLK-1), foetalliver kinase-4 (FLK-4) and the fms tyrosine kinase-1 (flt-1). The PDGFand FLK families are usually discussed together due to the similaritiesbetween the two groups. For a detailed discussion of receptor-typetyrosine kinases, see Plowman et al., DN & P 7(6): 334-339, 1994, whichis hereby incorporated by way of reference.

The non-receptor type tyrosine kinases likewise consist of amultiplicity of subfamilies, including Src, Frk, Btk, Csk, Abl, Zap70,Fes/Fps, Fak, Jak, Ack and L1MK. Each of these subfamilies is furthersub-divided into different receptors. For example, the Src subfamily isone of the largest subfamilies. It includes Src, Yes, Fyn, Lyn, Lck,Blk, Hck, Fgr and Yrk. The Src subfamily of enzymes has been linked tooncogenesis. For a more detailed discussion of non-receptor typetyrosine kinases, see Bolen Oncogene, 8: 2025-2031 (1993), which ishereby incorporated by way of reference.

Both receptor type tyrosine kinases and non-receptor type tyrosinekinases are involved in cellular signalling pathways leading to numerouspathogenic conditions, including cancer, psoriasis and hyperimmuneresponses. It has been proposed that various receptor-type tyrosinekinases, and the growth factors binding to them, play a role inangiogenesis, although some may promote angiogenesis indirectly(Mustonen and Alitalo, J. Cell Biol. 129: 895-898, 1995). One of thesereceptor-type tyrosine kinases is foetal liver kinase 1, also referredto as FLK-1. The human analogue of FLK-1 is the kinase insertdomain-containing receptor KDR, which is also known as vascularendothelial cell growth factor receptor 2 or VEGFR-2, since it bindsVEGF with high affinity. Finally, the murine version of this receptorhas also been called NYK (Oelrichs et al., Oncogene 8(1): 11-15, 1993).VEGF and KDR are a ligand-receptor pair which plays a vital role in theproliferation of vascular endothelial cells and the formation andsprouting of blood vessels, referred to as vasculogenesis andangiogenesis respectively. Angiogenesis is characterised by excessiveactivity of vascular endothelial growth factor (VEGF). VEGF actuallyconsists of a family of ligands (Klagsburn and D'Amore, Cytokine &Growth Factor Reviews 7: 259-270, 1996). VEGF binds the high affinitymembrane-spanning tyrosine kinase receptor KDR and the related fms-liketyrosine kinase-1, also known as Flt-1 or vascular endothelial cellgrowth factor receptor 1 (VEGFR-1). Cell culture and gene knockoutexperiments indicate that each receptor contributes to different aspectsof angiogenesis. KDR mediates the mitogenic function of VEGF, whereasFlt-1 appears to modulate non-mitogenic functions, such as thoseassociated with cellular adhesion. Inhibiting KDR thus modulates thelevel of mitogenic VEGF activity. In fact, tumour growth has been shownto be susceptible to the antiangiogenic effects of VEGF receptorantagonists (Kim et al., Nature 362, pp. 841-844, 1993).

Solid tumours can therefore be treated with tyrosine inhibitors sincethese tumours depend on angiogenesis for the formation of the bloodvessels that are necessary to support their growth. These solid tumoursinclude monocytic leukaemia, carcinomas of the brain, genito-urinarytract, lymphatic system, stomach, larynx and lung, including lungadenocarcinoma and small cell lung carcinoma. Further examples includecarcinomas in which overexpression or activation of Raf-activatingoncogenes (for example, K-Ras, Erb-B) is observed. Such carcinomasinclude pancreatic and breast carcinoma. Inhibitors of these tyrosinekinases are therefore suitable for the prevention and treatment ofproliferative diseases caused by these enzymes. The angiogenic activityof VEGF is not limited to tumours. VEGF accounts for the angiogenicactivity produced in or near the retina in diabetic retinopathy. Thisvascular growth in the retina leads to visual degeneration culminatingin blindness. Ocular VEGF mRNA and protein levels are elevated byconditions such as retinal vein occlusion in primates and decreased pO₂levels in mice that lead to neovascularization. Intraocular injectionsof anti-VEGF monoclonal antibodies or VEGF receptor immunofusionsinhibit ocular neovascularization in both primate and rodent models.Irrespective of the cause of induction of VEGF in human diabeticretinopathy, inhibition of ocular VEGF is suitable for treating thisdisease.

Expression of VEGF is also significantly increased in hypoxic regions ofanimal and human tumours adjacent to areas of necrosis. In addition,VEGF is upregulated by the expression of the oncogenes Ras, Raf, Src andmutant p53 (all of which are relevant in combating cancer). Anti-VEGFmonoclonal antibodies inhibit the growth of human tumours in nude mice.Although the same tumour cells continue to express VEGF in culture, theantibodies do not diminish their mitotic rate. Thus, tumour-derived VEGFdoes not function as an autocrine mitogenic factor. VEGF thereforecontributes to tumour growth in vivo by promoting angiogenesis throughits paracrine vascular endothelial cell chemotactic and mitogenicactivities. These monoclonal antibodies also inhibit the growth oftypically less well vascularised human colon carcinomas in athymic miceand decrease the number of tumours arising from inoculated cells.

The expression of a VEGF-binding construct of Flk-1, Flt-1, the mouseKDR receptor homologue truncated to eliminate the cytoplasmic tyrosinekinase domains but retaining a membrane anchor, virtually stops thegrowth of a transplantable glioblastoma in mice, presumably by thedominant negative mechanism of heterodimer formation withmembrane-spanning endothelial cell VEGF receptors.

Embryonic stem cells, which normally grow as solid tumours in nude mice,do not produce detectable tumours if both VEGF alleles are knocked out.Taken together, these data indicate the role of VEGF in the growth ofsolid tumours. Inhibition of KDR or Flt-1 is involved in pathologicalangiogenesis, and these receptors are suitable for the treatment ofdiseases in which angiogenesis is part of the overall pathology, forexample inflammation, diabetic retinal vascularization, as well asvarious forms of cancer, since tumour growth is known to be dependent onangiogenesis (Weidner et al., N. Engl. J. Med., 324, pp. 1-8, 1991).

Therefore, a preferred embodiment of the present invention is the use ofa compound of the present invention as VEGFR modulator or its use forthe prevention or treatment of VEGFR-mediated disorders.

Angiopoietin 1 (Ang1), a ligand for the endothelium-specificreceptor-type tyrosine kinase TIE-2, is a novel angiogenic factor (Daviset al., Cell, 1996, 87: 1161-1169; Partanen et al., Mol. Cell. Biol.,12: 1698-1707 (1992); U.S. Pat. Nos. 5,521,073; 5,879,672; 5,877,020;and 6,030,831). The acronym TIE stands for “tyrosine kinase with Ig andEGF homology domains”. TIE is used for the identification of a class ofreceptor-type tyrosine kinases, which are expressed exclusively invascular endothelial cells and early haemopoietic cells. TIE receptorkinases are typically characterised by the presence of an EGF-likedomain and an immunoglobulin (Ig)-like domain, which consists ofextracellular fold units stabilised by disulfide bridge bonds betweenthe chains (Partanen et al. Curr. Topics Microbiol. Immunol., 1999, 237:159-172). In contrast to VEGF, which exerts its function during theearly stages of vascular development, Ang1 and its receptor TIE-2 actduring the later stages of vascular development, i.e. during vasculartransformation (transformation relates to the formation of a vascularlumen) and maturing (Yancopoulos et al., Cell, 1998, 93:661-664; Peters,KG., Circ. Res., 1998, 83(3): 342-3; Suri et al., Cell 87, 1171-1180(1996)).

Accordingly, it would be expected that inhibition of TIE-2 shouldinterrupt the transformation and maturing of a new vascular systeminitiated by angiogenesis and should thus interrupt the angiogenesisprocess. Furthermore, inhibition at the kinase domain-binding site ofVEGFR-2 would block phosphorylation of tyrosine residues and serve tointerrupt initiation of angiogenesis. It must therefore be assumed thatinhibition of TIE-2 and/or VEGFR-2 should prevent tumour angiogenesisand serve to slow or completely eliminate tumour growth. Accordingly,treatment of cancer and other diseases associated with inappropriateangiogenesis could be provided.

The present invention also therefore relates to methods for theregulation, modulation or inhibition of TIE-2 for the prevention and/ortreatment of diseases associated with unregulated or disturbed TIE-2activity. In particular, the compounds according to the invention canalso be employed in the treatment of certain forms of cancer.Furthermore, the compounds according to the invention can be used toprovide additive or synergistic effects in certain existing cancerchemotherapies and/or can be used to restore the efficacy of certainexisting cancer chemotherapies and radio-therapies.

The protein kinase PKB (also known as AKT and RAC-PK) is a member of theAKT/PKB family of serine/threonine kinases and has been shown to beinvolved in a diverse set of signalling pathways in human malignancy(Nicholson et al., Cell. Signal., 2002, 14, 381-395). PKB, like othermembers of the AKT/PKB family, is located in the cytosol of unstimulatedcells and translocates to the cell membrane following stimulation. PKBtranslocation can be activated by a number of ligands, includingplatelet derived growth factor, epidermal growth factor, basicfibroblast growth factor, cellular stress, such as, for example, heatshock and hyperosmolarity, as well as insulin (Bos, Trends Biochem.Sci., 1995, 20, 441-442), and other studies have shown that thisactivation is through PI3 kinase which is wortmannin sensitive (Frankeet al., Science, 1997, 275, 665-668). Once localised to the plasmamembrane, PKB has been shown to mediate several functions within thecell, including apoptosis, the metabolic effects of insulin, inductionof differentiation and/or proliferation, protein synthesis and stressresponses (Alessi and Cohen, Curr. Opin. Genet. Dev., 1998, 8: 55-62;Downward, Curr. Opin. Cell Biol., 1998, 10, 262-267).

PKB was cloned independently in 1991 by three groups (Bellacosa et al.,Science, 1991, 254, 274-277; Coffer and Woodgett, Eur. J. Biochem.,1991, 201, 475-481; Jones et al., Cell Regul., 1991, 2, 1001-1009), butits association with primary human gastric carcinoma was recognised asearly as 1987 (Staal et al., Proc. Natl. Acad. Sci. USA, 1987, 84,5034-5037). Sequencing of PKBα revealed homology within the kinasedomains to the PKA (about 68%) and PKC isozymes (about 73%) (Jones etal., Proc. Natl. Acad. Sci. U.S.A., 1991, 88, 4171-5), a fact that leadto its renaming as PKB. There are three cellular isoforms of PKB and twosplice variants (PKBα, β, γ, β₁, γ₁; Brazil et al. Trends in Bio Sci,2001, 26, 657-663). PKBα was found to be amplified or overexpressed ingastric adenocarcinomas and in a breast cancer cell line (Staal et al.,Proc. Natl. Acad. Sci. U.S.A., 1987, 84, 5034-7; Jones et al., CellRegul., 1991, 2, 1001-9). PKBβ is amplified or overexpressed in 3% ofbreast (Bellacosa et al., Int. J. Cancer, 1995 64, 280-5), 12% ofpancreatic (Cheng et al., Proc. Natl. Acad. Sol. U.S.A., 1996, 93,3636-41) and 15% of ovarian cancers (Bellacosa et al., Int. J. Cancer,1995, 64, 280-5; Cheng et al., Proc. Natl. Acad. Sci. U.S.A., 1992, 89,9267-71).

PKBγ is overexpressed in oestrogen receptor-deficient breast cancer andin androgen-independent prostate cell lines (Nakatani et al., J. Biol.Chem. 1999, 274, 21528-32). It has been proposed that PKB is a gene,which is involved in chromosomal rearrangement at chromosome band 1402.This locus is known to undergo rearrangement in human T-cellmalignancies, such as, for example, prolymphocytic leukaemias and mixedlineage childhood leukaemias (Staal et al., Genomics, 1988, 2, 96-98).

PKB also plays a role in the prevention of “programmed cell death” orapoptosis by inhibitory phosphorylation of ASK-1, Bad, Caspase9 and FKHR(for review see Nicholson et al., Cell Signalling 2001, 14, 281-395). Ithas been shown that PKB provides a survival signal (for review seeLawlor et al., J. of Cell Science 2001, 114, 2903-2910) to cells inorder to protect them from a number of agents, including UV radiation(Dudek et al., Science, 1997, 275, 661-665), withdrawal of IGF1 fromneuronal cells, detachment from the extracellular matrix, stress andheat shock (Alessi and Cohen, Curr. Opin. Genet. Dev., 1998, 8: 55-62).

The dual-specific phosphatase PTEN (phosphatase and tensin homologuedeleted on chromosome ten) increases the Ptdlns(3, 4, 5)P₃ level in thecell by dephosphorylation of Ptdlns(3, 4, 5)P₃. Ptdlns(3, 4, 5)P₃ bindsto the PH domain (Pleckstrin homology domain) of PKB. This binding is anessential step for membrane translocation and activation of PKB. PTEN isa tumour suppressor gene mutated in a large proportion of glioblastomaand melanoma cell lines, advanced prostate carcinomas and endometrialcarcinomas. Furthermore, it is deleted in >80% of patients withhereditary conditions, such as, for example, Cowden's disease,Lhermitte-Duclose disease and Bannayan-Zonana Syndrome. The patientsdisplay a number of similar features, including multiple benign tumours(harmatomas) and increased susceptibility to breast and thyroidmalignancies (Di Cristofano et al. Cell, 2000, 100, 387-390).

Cell lines derived from PTEN^(+/−) heterozygous mice (PTEN^(−/−)heterozygous mice are not viable) show increased Ptdlns(3, 4, 5)P₃levels paralleled by increased PKB activity, with concomitant decreasedsensitivity to apoptosis (Di Christofano et al. Nat. Genet. 1998, 19,348-355; Stambolic et al., Cell, 1998, 95, 29-39, Myers et al., Proc.Natl. Acad. Si. U.S.A., 1998, 96: 13513-13518). PKB is also able topromote cell cycle progression by inhibiting p21 cell cycle inhibitor(Zhou et al.; Nat. Cell Biol., 2002, 3: 245-252). These findings mayexplain the overexpression of PKB observed in cancer cells, which allowspreferential survival and proliferation of the carcinomas by avoidingthe normal progression to apoptosis.

Therefore, a preferred embodiment of the present invention is the use ofa compound of the present invention as PKB modulator or its use for theprevention or treatment of PKB-mediated disorders.

Some of the most common mutations leading to upregulation of mitogenicsignals and proliferative diseases occur in genes encoding the proteinknown as Ras, a G-protein that is activated when bound to GTP, andinactivated when bound to GDP. The above-mentioned growth factorreceptors, and many other mitogenic receptors, when activated, lead toRas being converted from the GDP-bound state to the GTP-bound state.This signal is an absolute prerequisite for proliferation in most celltypes. Defects in this signaling system, especially in the deactivationof the Ras-GTP complex, are common in cancers, and lead to the signalingcascade below Ras being chronically activated.

Activated Ras leads in turn to the activation of a cascade ofserine/threonine kinases. One of the groups of kinases known to requirean active Ras-GTP for its own activation is the Raf family.

The present invention therefore relates to the compounds of the presentinvention as inhibitors of Raf kinases. Protein phosphorylation is afundamental process for the regulation of cellular functions. Thecoordinated action of both protein kinases and phosphatases controls thedegrees of phosphorylation and, hence, the activity of specific targetproteins. One of the predominant roles of protein phosphorylation is insignal transduction, where extracellular signals are amplified andpropagated by a cascade of protein phosphorylation and dephosphorylationevents, for example in the p21^(ras)/Raf pathway.

The p21^(ras) gene was discovered as an oncogene of the Harvey (H-Ras)and Kirsten (K-Ras) rat sarcoma viruses. In humans, characteristicmutations in the cellular Ras gene (c-Ras) have been associated withmany different types of cancers. These mutant alleles, which render Rasconstitutively active, have been shown to transform cells, such, forexample, as the murine cell line NIH 3T3, in culture.

The p21^(ras) oncogene is a major contributor to the development andpro-gression of human solid carcinomas and is mutated in 30% of allhuman carcinomas (Bolton et al. (1994) Ann. Rep. Med. Chem., 29, 165-74;Bos. (1989) Cancer Res., 49: 4682-9). In its normal, unmutated form, theRas protein is a key element of the signal transduction cascade directedby growth factor receptors in almost all tissues (Avruch et al. (1994)Trends Biochem. Sci., 19: 279-83).

Biochemically, Ras is a guanine nucleotide binding protein and cyclingbetween a GTP-bound activated and a GDP-bound resting form is strictlycontrolled by Ras endogenous GTPase activity and other regulatoryproteins. The Ras gene product binds to guanine triphosphate (GTP) andguanine diphosphate (GDP) and hydrolyses GTP to GDP. Ras is active inthe GTP-bound state. In the Ras mutants in cancer cells, the endogenousGTPase activity is reduced and the protein consequently transmitsconstitutive growth signals to downstream effectors, such as, forexample, the enzyme Raf kinase. This leads to the cancerous growth ofthe cells, which carry these mutants (Magnuson et al. (1994) Semin.Cancer Biol., 5, 247-53). The Ras proto-oncogene requires a functionallyintact c-Raf-1 proto-oncogene in order to transduce growth anddifferentiation signals initiated by receptor and non-receptor-typetyrosine kinases in higher eukaryotes.

Activated Ras is necessary for the activation of the c-Raf-1proto-oncogene, but the biochemical steps through which Ras activatesthe Raf-1 protein (Ser/Thr) kinase are now well characterised. It hasbeen shown that inhibiting the effect of active Ras by inhibiting theRaf kinase signalling pathway by administration of deactivatingantibodies to Raf kinase or by co-expression of dominant negative Rafkinase or dominant negative MEK (MAPKK), the substrate of Raf kinase,leads to reversion of transformed cells to the normal growth phenotype(see: Daum et al. (1994) Trends Biochem. Sci., 19, 474-80; Fridman etal. (1994) J. Biol. Chem., 269, 30105-8. Kolch et al. (1991) Nature,349, 426-28 and for a review Weinstein-Oppenheimer et al. Pharm. &Therap. (2000), 88, 229-279).

Similarly, inhibition of Raf kinase (by antisense oligodeoxynucleotides)has been correlated in vitro and in vivo with inhibition of the growthof a variety of human tumour types (Monia et al., Nat. Med. 1996, 2,668-75).

Raf serine- and threonine-specific protein kinases are cytosolic enzymesthat stimulate cell growth in a variety of cell systems (Rapp, U. R., etal. (1988) in The Oncogene Handbook; T. Curran, E. P. Reddy and A.Skalka (eds.) Elsevier Science Publishers; The Netherlands, pp. 213-253;Rapp, U. R., et al. (1988) Cold Spring Harbor Sym. Quant. Biol. 53:173-184; Rapp, U. R., et al. (1990) Inv Curr. Top. Microbial. Immunol.Potter and Melchers (eds.), Berlin, Springer-Verlag 166: 129-139).

Three isozymes have been characterised: C-Raf (Raf-1) (Bonner, T. I., etal. (1986) Nucleic Acids Res. 14: 1009-1015). A-Raf (Beck, T. W., et al.(1987) Nucleic Acids Res. 15: 595-609) and B-Raf (Qkawa, S., et al.(1998) Mol. Cell. Biol. 8: 2651-2654; Sithanandam, G. et al. (1990)Oncogene: 1775). These enzymes differ in their expression in varioustissues. Raf-1 is expressed in all organs and in all cell lines thathave been examined, and A- and B-Raf are expressed in urogenital andbrain tissues respectively (Storm, S. M. (1990) Oncogene 5: 345-351).

Raf genes are proto-oncogenes: they can initiate malignanttransformation of cells when expressed in specifically altered forms.Genetic changes that lead to oncogenic activation generate aconstitutively active protein kinase by removal of or interference withan N-terminal negative regulatory domain of the protein (Heidecker, G.,et al. (1990) Mol. Cell. Biol. 10: 2503-2512; Rapp, U. R., et al. (1987)in Oncogenes and Cancer; S. A. Aaronson, J. Bishop, T. Sugimura, M.Terada, K. Toyoshima and P. K. Vogt (eds.) Japan Scientific Press,Tokyo). Microinjection into NIH 3T3 cells of oncogenically activated,but not wild-type, versions of the Raf protein prepared with Escherichiacoli expression vectors results in morphological transformation andstimulates DNA synthesis (Rapp, U. R., et al. (1987) in Oncogenes andCancer; S. A. Aaronson, J. Bishop, T. Sugimura, M. Terada, K. Toyoshimaand P. K. Vogt (ed.) Japan Scientific Press, Tokyo; Smith, M. R., et al.(1990) Mol. Cell. Biol. 10: 3828-3833).

Consequently, activated Raf-1 is an intracellular activator of cellgrowth. Raf-1 protein serine kinase is a candidate for the downstreameffector of mitogen signal transduction, since Raf oncogenes overcomegrowth arrest resulting from a block of cellular Ras activity due eitherto a cellular mutation (Ras revertant cells) or microinjection ofanti-Ras antibodies (Rapp, U. R., et al. (1988) in The OncogeneHandbook, T. Curran, E. P. Reddy and A. Skalka (ed.), Elsevier SciencePublishers; The Netherlands, pp. 213-253; Smith, M. R., et al. (1986)Nature (London) 320: 540-543).

C-Raf function is required for transformation by a variety ofmembrane-bound oncogenes and for growth stimulation by mitogenscontained in serums (Smith, M. R., et al. (1986) Nature (London) 320:540-543). Raf-1 protein serine kinase activity is regulated by mitogensvia phosphorylation (Morrison, D. K., et al. (1989) Cell 58: 648-657),which also effects sub-cellular distribution (Olah, Z., et al. (1991)Exp. Brain Res. 84: 403; Rapp, U. R., et al. (1988) Cold Spring HarborSym. Quant. Biol. 53: 173-184. Raf-1 activating growth factors includeplatelet-derived growth factor (PDGF) (Morrison, D. K., et al. (1988)Proc. Natl. Acad. Sci. USA 85: 8855-8859), colony-stimulating factor(Baccarini, M., et al. (1990) EMBO J. 9: 3649-3657), insulin(Blackshear, P. J., et al. (1990) J. Biol. Chem. 265: 12115-12118),epidermal growth factor (EGF) (Morrison, R. K., et al. (1988) Proc.Natl. Acad. Sci. USA 85: 8855-8859), interleukin-2 (Turner, B. C., etal. (1991) Proc. Natl. Acad. Sci. USA 88:1227) and interleukin-3 andgranulocyte macrophage colony-stimulating factor (Carroll, M. P., et al.(1990) J. Biol. Chem. 265: 19812-19817).

After mitogen treatment of cells, the transiently activated Raf-1protein serine kinase translocates to the perinuclear area and thenucleus (Olah, Z., et al. (1991) Exp. Brain Res. 84: 403; Rapp, U. R.,et al. (1988) Cold Spring Harbor Sym. Quant. Biol. 53: 173-184). Cellscontaining activated Raf are altered in their pattern of gene expression(Heidecker, G., et al. (1989) in Genes and signal transduction inmultistage carcinogenesis, N. Colburn (ed.), Marcel Dekker, Inc., NewYork, pp. 339-374) and Raf oncogenes activate transcription fromAp-I/PEA3-dependent promoters in transient transfection assays (Jamal,S., et al. (1990) Science 344: 463-466; Kaibuchi, K., et al. (1989) J.Biol. Chem. 264: 20855-20858; Wasylyk, C., et al. (1989) Mol. Cell.Biol. 9: 2247-2250).

There are at least two independent pathways for Raf-1 activation byextracellular mitogens: one involving protein kinase C (KC) and a secondinitiated by protein tyrosine kinases (Blackshear, P. J., et al. (1990)J. Biol. Chem. 265:12131-12134; Kovacina, K. S., et al. (1990) J. Biol.Chem. 265: 12115-12118; Morrison, D. K., et al. (1988) Proc. Natl. Acad.Sci. USA 85: 8855-8859; Siegel, J. N., et al. (1990) J. Biol. Chem. 265:18472-18480; Turner, B. C., et al. (1991) Proc. Natl. Acad. Sci. USA 88:1227). In each case, activation involves Raf-1 protein phosphorylation.Raf-1 phosphorylation may be a consequence of a kinase cascade amplifiedby autophosphorylation or may be caused entirely by autophosphorylationinitiated by binding of a putative activating ligand to the Raf-1regulatory domain, analogous to PKC activation by diacylglycerol(Nishizuka, Y. (1986) Science 233: 305-312).

MAPK/ERK Kinase (“MEK”) enzymes are dual specificity kinases involvedin, for example, immunomodulation, inflammation, and proliferativediseases such as cancer and restenosis.

These kinases of the Raf family activate MEK (e.g., MEK1 and MEK2) whichthen activates the MAP kinase, ERK (ERK1 and ERK2). Activation of MAPkinase by mitogens appears to be essential for proliferation,constitutive activation of this kinase is sufficient to induce cellulartransformation. Blockade of downstream Ras signaling, for example by useof a dominant negative Raf-1 protein, can completely inhibitmitogenesis, whether induced from cell surface receptors or fromoncogenic Ras mutants. Although Ras is not itself a protein kinase, itparticipates in the activation of Raf and other kinases, most likelythrough a phosphorylation mechanism. Once activated, Raf and otherkinases phosphorylate MEK on two closely adjacent serine residues, S²¹⁸and S²²² in the case of MEK1, which are the prerequisite for activationof MEK as a kinase. MEK in turn phosphorylates MAP kinase on both atyrosine, Y¹⁸⁵, and a threonine residue, T¹⁸³, separated by a singleamino acid. This double phosphorylation activates MAP kinase at least100-fold. Activated MAP kinase can then catalyze the phosphorylation ofa large number of proteins, including several transcription factors andother kinases. Many of these MAP kinase phosphorylations aremitogenically activating for the target protein, such as a kinase, atranscription factor, or another cellular protein. In addition to Raf-1and MEKK, other kinases activate MEK, and MEK itself appears to be asignal integrating kinase. Current understanding is that MEK is highlyspecific for the phosphorylation of MAP kinase. In fact, no substratefor MEK other than the MAP kinase, ERK, has been demonstrated to dateand MEK does not phosphorylate peptides based on the MAP kinasephosphorylation sequence, or even phosphorylate denatured MAP kinase.MEK also appears to associate strongly with MAP kinase prior tophosphorylating it, suggesting that phosphorylation of MAP kinase by MEKmay require a prior strong interaction between the two proteins. Boththis requirement and the unusual specificity of MEK are suggestive thatit may have enough difference in its mechanism of action to otherprotein kinases that selective inhibitors of MEK, possibly operatingthrough allosteric mechanisms rather than through the usual blockade ofthe ATP binding site, may be found.

It has been found that the compounds of the present invention areinhibitors of MEK and are useful in the treatment of a variety ofproliferative disease states, such as conditions related to thehyperactivity of MEK, as well as diseases modulated by the MEK cascade.

Selective MEK1 or MEK2 inhibitors are those compounds, which inhibit theMEK1 or MEK2 enzymes, respectively, without substantially inhibitingother enzymes such as MKK3, PKC, Cdk2A, phosphorylase kinase, EGF andC-src. In general, a selective MEK1 or MEK2 inhibitor has an IC₅₀ forMEK1 or MEK2 that is at least one-fiftieth ( 1/50) that of its IC₅₀ forone of the above-named other enzymes. Preferably, a selective inhibitorhas an IC₅₀ that is at least 1/100, more preferably 1/500, and even morepreferably 1/1000, 1/5000, or less than that of its IC₅₀ or one or moreof the above-named enzymes.

The identification of small compounds, which specifically inhibit,regulate and/or modulate signal transduction of protein kinases and canbe used as medicaments for the treatment of various diseases istherefore desirable and an aim of the present invention.

The disclosed compounds are useful as both prophylactic and therapeutictreatments for disorders or conditions related to the change of activityof one or more of the above-mentioned kinases, especially thehyperactivity of MEK, as well as diseases or conditions modulated bysaid kinase cascades.

Thus, a further preferred embodiment of the present invention is the useof a compound of the formula I for the preparation of a medicament forthe treatment and/or prevention of disorders.

The invention therefore also relates to compounds of the formula I andthe physiologically acceptable salts, derivatives, prodrugs, solvatesand stereoisomers thereof, including mixtures thereof in all ratios asmedicaments.

A further preferred embodiment of the present invention is the use ofthe compounds according to the invention for the manufacture of amedicament for the treatment and/or prevention of disorders, which arecaused, mediated and/or propagated by protein kinases.

A further preferred embodiment of the present invention is the use ofthe compounds according to the invention for the manufacture of amedicament for the treatment and/or prevention of disorders, which arecaused, mediated and/or propagated by protein kinases, characterized inthat the protein kinases are MEK1 or MEK2.

Usually, the disorders discussed herein are divided into two groups,hyperproliferative and non-hyperproliferative disorders. In thiscontext, infection or infectious diseases, psoriasis, arthritis,inflammation, endometriosis, scarring, begnin prostatic hyperplasia,immunological diseases, autoimmune diseases and immunodeficiencydiseases are to be regarded as non-cancerous disorders, of whichinfection, arthritis, inflammation, immunological diseases, autoimmunediseases and immunodeficiency diseases are usually regarded asnon-hyperproliferative disorders. In this context, brain cancer, lungcancer, squamous cell cancer, bladder cancer, gastric cancer, pancreaticcancer, hepatic cancer, renal cancer, colorectal cancer, breast cancer,head cancer, neck cancer, oesophageal cancer, gynaecological cancer,thyroid cancer, lymphoma, chronic leukaemia and acute leukaemia are tobe regarded as cancerous disorders, all of which are usually regarded ashyperproliferative disorders.

Thus, a preferred embodiment of the present invention is the use of acompound of the formula I for the preparation of a medicament for thetreatment and/or prevention of disorders, characterized in that thedisorders are selected from the group consisting of hyperproliferativeand non-hyperproliferative disorders.

In a preferred embodiment of the present invention the disorder isnon-cancerous.

Therefore, the compounds of the formula I can be used for thepreparation of a medicament for the treatment and/or prevention ofdisorders, which are selected from the group consisting of psoriasis,arthritis, rheumatoid arthritis, inflammation, endometriosis, scarring,Helicobacter pylori Influenza A infection, HIV infection, hepatitis (B)virus (HBV) infection, human papilloma virus (HPV) infection,cytomegalovirus (CMV) infection, and Epstein-Barr virus (EBV) infection,begnin prostatic hyperplasia, immunodeficiency diseases, autoimmunedisease, immunological diseases, chronic obstructive pulmonary disease,asthma, inflammatory bowel disease, fibrosis, atherosclerosis,restenosis, vascular disease, cardiovascular disease, stroke (such asacute focal ischemic stroke and global cerebral ischemia), heartfailure, cystic fibrosis, hepatomegaly, cardiomegaly, septic shock,Alzheimer's disease, chronic or neuropathic pain, renal disease andangiogenesis disorders, mesangial cell proliferative disorders, diabeticnephropathy, diabetic retinopathy, malignant nephrosclerosis, thromboticmicroangiopathy syndromes, xenograft (cell(s), skin, limb, organ or bonemarrow transplant) rejection, glomerulopathies, metabolic disorders andneurodegenerative diseases.

Infections according the invention include, but are not limited toinfections caused by pathogenic microorganisms, such as bacteria, fungi,viruses and protozoans, for example influenza (Pleschka, S. et al.Nature Cell Biol. 2001, 3, page 301-305), retroviruses, for example HIVinfection (Yang, X. et al. J. Biol. Chem. 1999, 274, page 27981-27988;Popik, W et al Mol Cell Biol. 1996, 16, page 6532-6541), Hepatitis B(Benn, J et al., Proc. Natl. Acad. Sci. 1995, 92, page 11215-11219),Hepatitis C (Aoki et al. J. Virol. 2000, 74, page 1736-1741),papillomavirus, parainfluenza, rhinoviruses, adenoviruses, Heliobacterpylori, and viral and bacterial infections of the skin (e.g. cold sores,warts, chickenpox, molluscum, contagiosum, herpes zoster, boils,cellulitis, erysipelas, impetigo, tinea, Althlete's foot and ringworm).

Furthermore, a preferred embodiment of the present invention is the useof a compound of the formula I for the manufacture of a medicament forthe treatment and/or prevention of disorders, characterized in that thedisorders are selected from the group consisting of hyperproliferativedisorders.

There are many disorders associated with a dysregulation of cellularproliferation. The conditions of interest include, but are not limitedto, the following conditions. The subject compounds are useful in thetreatment of a variety of conditions where there is proliferation and/ormigration of smooth muscle cells, and/or inflammatory cells into theintimal layer of a vessel, resulting in restricted blood flow throughthat vessel, e.g., neointimal occlusive lesions. Occlusive vascularconditions of interest include atherosclerosis, graft coronary vasculardisease after transplantation, vein graft stenosis, peri-anastomaticprothetic graft stenosis, restenosis after angioplasty or stentplacement, and the like.

Furthermore, the compounds of the formula I preferably showanti-angiogenic properties. Thus, compounds of the present invention canbe advantageously employed in the treatment of one or more diseasesafflicting mammals which are characterized by cellular proliferation inthe area of disorders associated with neo-vascularization and/orvascular permeability including blood vessel proliferative disordersincluding arthritis and restenosis; fibrotic disorders including hepaticcirrhosis and atherosclerosis; mesangial cell proliferative disordersinclude glomerulonephritis, diabetic nephropathy, malignantnephrosclerosis, thrombotic microangiopathy syndromes, organ transplantrejection and glomerulopathies; and metabolic disorders includepsoriasis, diabetes mellitus, chronic wound healing, inflammation andneurodegenerative diseases.

The process of angiogenesis is the development of new blood vessels,generally capillaries, from pre-existing vasculature. Angiogenesis isdefined as involving (i) activation of endothelial cells; (ii) increasedvascular permeability; (iii) subsequent dissolution of the basementmembrane and extravisation of plasma components leading to formation ofa provisional fibrin gel extracellular matrix; (iv) proliferation andmobilization of endothelial cells; (v) reorganization of mobilizedendothelial cells to form functional capillaries; (vi) capillary loopformation; and (vii) deposition of basement membrane and recruitment ofperivascular cells to newly formed vessels. Normal angiogenesis isactivated during tissue growth, from embryonic development throughmaturity, and then enters a period of relative quiescence duringadulthood.

Normal angiogenesis is also activated during wound healing, and atcertain stages of the female reproductive cycle. Inappropriate orpathological angiogenesis has been associated with several diseasestates including various retinopathies; ischemic disease;atherosclerosis; chronic inflammatory disorders; rheumatoid arthritis,and cancer. The role of angiogenesis in disease states is discussed, forinstance, in Fan et al., Trends in Pharmacol Sci. 16:54 66; Shawver etal., DOT Vol. 2, No. 2 Feb. 1997; Folkmann, 1995, Nature Medicine1:27-31.

In cancer the growth of solid tumors has been shown to be angiogenesisdependent. (see Folkmann, J., J. Nat'l. Cancer Inst., 1990, 82, 4-6.)Consequently, the targeting of pro-angiogenic pathways is a strategybeing widely pursued in order to provide new therapeutics in these areasof great, unmet medical need.

Several protein kinases are involved in angiogenic processes.Endothelial growth factors (e.g. vascular endothelial growth factorVEGF) activate receptor tyrosine kinases (e.g. VEGFR-2) and signalthrough the Ras/Raf/Mek/Erk kinase cascade. Activation of VEGFR-2 byVEGF is a critical step in the signal transduction pathway thatinitiates tumor angiogenesis. VEGF expression may be constitutive totumor cells and can also be upregulated in response to certain stimuli.One such stimuli is hypoxia, where VEGF expression is upregulated inboth tumor and associated host tissues. The VEGF ligand activatesVEGFR-2 by binding with its extracellular VEGF binding site. This leadsto receptor dimerization of VEGFRs and autophosphorylation of tyrosineresidues at the intracellular kinase domain of VEGFR-2. The kinasedomain operates to transfer a phosphate from ATP to the tyrosineresidues, thus providing binding sites for signaling proteins downstreamof VEGFR-2 leading ultimately to initiation of angiogenesis (McMahon,G., The Oncologist, Vol. 5, No. 90001, 3-10, April 2000).

Mice with a targeted disruption in the B-raf gene die of vasculardefects during development (Wojnowski, L. et al. 1997, Nature genetics16, page 293-296). These mice show defects in the formation of thevascular system and in angiogenesis e.g. enlarged blood vessels andincreased apoptotic death of differentiated endothelial cells.

Diseases where there is hyperproliferation and tissue remodelling orrepair or reproductive tissue, e.g., uterine, testicular and ovariancarcinomas, endometriosis, squamous and glandular epithelial carcinomasof the cervix, etc. are reduced in cell number by administration of thesubject compounds. The growth and proliferation of neural cells is alsoof interest.

Tumor cells are characterized by uncontrolled growth, invasion tosurrounding tissues, and metastatic spread to distant sites. Growth andexpansion requires an ability not only to proliferate, but also todown-modulate cell death (apoptosis) and activate angiogenesis toproduce a tumor neovasculature.

A preferred embodiment of the present invention is the use of a compoundof the formula I for the manufacture of a medicament for the treatmentand/or prevention of disorders, characterized in that the disorder iscancer, whether solid or hematopoietic.

Therefore, a preferred embodiment of the present invention is the use ofa compound of the general formula I for the manufacture of a medicamentfor the treatment and/or prevention of disorders, characterised in thatthe disorders are selected from the group consisting of carcinomas, e.g.melanoma, brain cancer, lung cancer, non-small cell lung carcinoma,transitional and squamous cell urinary carcinoma, bladder cancer,gastric cancer, pancreatic cancer, colon cancer, duodenal cancer, ductalcancer, endometrial cancer, stomach cancer, colorectal cancer, hepaticcancer, renal cancer, breast cancer, head cancer, neck cancer,oesophageal cancer, gynaecological cancer, ovarian cancer, uterinecancer, prostate cancer, thyroid cancer, dysplastic oral mucosa,polyposis, invasive oral cancer, etc.; neurological malignancies; e.g.neuroplastoma, gliomas, etc; hematological malignancies, e.g., childhoodacute leukaemia, non-Hodgkin's lymphomas, chronic lymphocytic leukaemia,malignant cutaneous T-cells, mycosis fungoides, non-MF cutaneousT-cell-lymphoma, lymphomatoid papulosis, T-cell rich cutaneous lymphoidhyperplasia, bullous pemphigoid, discoid lupus erythematosus, lichenplanus, etc.; and the like.

Tumors of neural tissue are of particular interest, e.g., gliomas,neuromas, etc. Some cancers of particular interest include breastcancers, which are primarily adenocarcinoma subtypes. Ductal carcinomain situ is the most common type of noninvasive breast cancer. In DCIS,the malignant cells have not metastasized through the walls of the ductsinto the fatty tissue of the breast. Infiltration (or invasive) ductalcarcinoma (IDC) has metastasized through the wall of the duct andinvaded the fatty tissue of the breast. Infiltrating (or invasive)lobular carcinoma (ILC) is similar to IDC, in that it has the potentialto metastasize elsewhere in the body. About 10% to 15% of invasivebreast cancers are invasive lobular carcinomas.

Also of interest is non-small cell lung carcinoma. Non-small cell lungcancer (NSCLC) is made up of three general subtypes of lung cancer.Epidermoid carcinoma (also called squamos cell carcinoma) usually startsin one of the larger bronchial tubes and grows relatively slowly. Thesize of these tumors can range from very small to quite large.Adenocarcinoma starts growing near the outside surface of the lung andmay vary in both size and growth rate. Some slowly growingadenocarcinomas are described as alveolar cell cancer. Large cellcarcinoma starts near the surface of the lung, grows rapidly, and thegrowth is usually fairly large when diagnosed. Other less common formsof lung cancer are carcinoid, cylindroma, mucoepidermoid, and malignantmesothelioma.

Melanoma is a malignant tumor of melanocytes. Although most melanomasarise in the skin, they also may arise from mucosal surfaces or at othersites to which neural crest cells migrate. Melanoma occurs predominantlyin adults, and more than half of the cases arise in apparently normalareas of the skin. Prognosis is affected by clinical and histologicalfactors and by anatomic location of the lesion. Thickness and/or levelof invasion of the melanoma, mitotic index, tumor infiltratinglymphocytes, and ulceration or bleeding at the primary site affect theprognosis. Clinical staging is based on whether the tumor has spread toregional lymph nodes or distant sites. For disease, clinically confinedto the primary site, the greater the thickness and depth of localinvasion of the melanoma, the higher the chance of lymph node metastasesand the worse the prognosis. Melanoma can spread by local extension(through lymphatics) and/or by hematogenous routes to distant sites. Anyorgan may be involved by metastases, but lungs and liver are commonsites.

Other hyperproliferative diseases of interest relate to epidermalhyperproliferation, tissue, remodeling and repair. For example, thechronic skin inflammation of psoriasis is associated with hyperplasticepidermal keratinocycles as well as infiltrating mononuclear cells,including CD4+ memory T cells, neutrophils and macrophages.

The proliferation of immune cells is associated with a number ofautoimmune and lymphoproliferative disorders. Diseases of interestinclude multiple sclerosis, rheumatoid arthritis and insulin dependentdiabetes mellitus. Evidence suggests that abnormalities in apoptosisplay a part in the pathogenesis of systemic lupus erythematosus (SLE).Other lymphoproliferative conditions the inherited disorder oflymphocyte apoptosis, which is an autoimmune lymphoproliferativesyndrome, as well as a number of leukemia's and lymphomas. Symptoms ofallergies to environmental and food agents, as well as inflammatorybowel disease, may also be alleviated by the compounds of the invention.

For use in the subject methods, the subject compounds may be formulatedwith pharmaceutically active agents other than the compounds accordingto the invention, particularly other anti-metastatic, antitumor oranti-angiogenic agents. Angiostatic compounds of interest includeangiostatin, enclostatin, carboxy terminal peptides of collagen alpha(XV), etc. Cytotoxic and cytostatic agents of interest includeadriamycin, aleran, Ara-C, BICNU, busulfan, CNNU, cisplatinum, cytoxan,daunorubicin, DTIC, 5-FU, hydrea, ifosfamicle, methotrexate,mithramycin, mitomycin, mitoxantrone, nitrogen mustard, velban,vincristine, vinblastine, VP-16, carboplatinum, fludarabine,gemcitabine, idarubicin, irinotecan, leustatin, navelbine, taxol,taxotere, topotecan, etc. For example, in the case of bone conditions,combinations that would be favourable include those with antiresorptivebisphosphonates, such as alendronate and risedronate; integrin blockers(as defined further below), such as α_(v)β₃ antagonists; conjugatedoestrogens used in hormone replacement therapy, such as PREMPRO®,PREMARIN® and ENDOMETRION®; selective oestrogen receptor modulators(SERMs), such as raloxifene, droloxifene, CP-336,156 (Pfizer) andlasofoxifene; cathepsin K inhibitors; and ATP proton pump inhibitors.

The present compounds are also suitable for combination with knownanti-cancer agents. These known anti-cancer agents include thefollowing: oestrogen receptor modulators androgen receptor modulators,retinoid receptor modulators, cytotoxic agents, antiproliferativeagents, prenyl-protein transferase inhibitors, HMG-CoA reductaseinhibitors, HIV protease inhibitors, reverse transcriptase inhibitorsand other angiogenesis inhibitors. The present compounds areparticularly suitable for administration at the same time asradiotherapy. The synergistic effects of inhibiting VEGF in combinationwith radiotherapy have been described in the art (see WO00/61186).

Therefore, a preferred embodiment of the present invention is the use ofa compound of the general formula I for the manufacture of a medicamentfor the treatment and/or prevention of disorders, characterized in thata therapeutically effective amount of one or more compounds according tothe present invention is administered in combination with an compoundselected from the group consisting of estrogen receptor modulators,androgen receptor modulators, retinoid receptor modulators, cytotoxicagents, anti-proliferative agents, prenyl protein protease inhibitors,HMG CoA reductase inhibitors, HIV protease inhibitors, reversetranscriptase inhibitors, growth factor receptor inhibitors andangiogenesis inhibitors.

Additionally, a preferred embodiment of the present invention is the useof a compound of the general formula I for the manufacture of amedicament for the treatment and/or prevention of disorders,characterized in that a therapeutically effective amount of one or morecompounds according to the present invention is administered incombination radio therapy and with an compound selected from the groupconsisting of estrogen receptor modulators, androgen receptormodulators, retinoid receptor modulators, cytotoxic agents,anti-proliferative agents, prenyl protein protease inhibitors, HMG CoAreductase inhibitors, HIV protease inhibitors, reverse transcriptaseinhibitors, growth factor receptor inhibitors and angiogenesisinhibitors.

“Oestrogen receptor modulators” refers to compounds, which interferewith or inhibit the binding of oestrogen to the receptor, regardless ofmechanism. Examples of oestrogen receptor modulators include, forexample, tamoxifen, raloxifene, idoxifene, LY353381, LY 117081,toremifene, fulvestrant,4-[7-(2,2-dimethyl-1-oxopropoxy-4-methyl-2-[4-[2-(1-piperidinyl)ethoxy]phenyl]-2H-1-benzopyran-3-yl]phenyl-2,2-dimethylpropanoate,4,4″-dihydroxybenzophenone-2,4-dinitrophenylhydrazone and SH646.“Androgen receptor modulators” refers to compounds, which interfere withor inhibit the binding of androgens to the receptor, regardless ofmechanism. Examples of androgen receptor modulators include finasterideand other 5α-reductase inhibitors, nilutamide, flutamide, bicalutamide,liarozole and abiraterone acetate.

“Retinoid receptor modulators” refers to compounds, which interfere withor inhibit the binding of retinoids to the receptor, regardless ofmechanism. Examples of such retinoid receptor modulators includebexarotene, tretinoin, 13-cis-retinoic acid, 9-cis-retinoic acid,α-difluoromethylornithine, ILX23-7553,trans-N-(4′-hydroxyphenyl)retinamide and N-4-carboxyphenyl retinamide.

“Cytotoxic agents” refers to compounds, which result in cell deathprimarily through direct action on the cellular function or inhibit orinterfere with cell myosis, including alkylating agents, tumour necrosisfactors, intercalators, microtubulin inhibitors and topoisomeraseinhibitors.

Examples of cytotoxic agents include, but are not limited to,tirapazimine, sertenef, cachectin, ifosfamide, tasonermin, lonidamine,carboplatin, altretamine, prednimustine, dibromodulcitol, ranimustine,fotemustine, nedaplatin, oxaliplatin, temozolomide, heptaplatin,estramustine, improsulfan tosylate, trofosfamide, nimustine,dibrospidium chloride, pumitepa, lobaplatin, satraplatin, profiromycin,cisplatin, irofulven, dexifosfamide, cis-aminedichloro(2-methylpyridine)platinum, benzylguanine, glufosfamide, GPX100,(trans,trans,trans)-bis-mu-(hexane-1,6-diamine)-mu-[diamine-platinum(II)]bis[diamine(chloro)platinum(II)]tetrachloride, diarizidinyl-spermine, arsenic trioxide,1-(11-dodecylamino-10-hydroxyundecyl)-3,7-dimethylxanthine, zorubicin,idarubicin, daunorubicin, bisantrene, mitoxan-trone, pirarubicin,pinafide, valrubicin, amrubicin, antineoplaston,3′-deamino-3′-morpholino-13-deoxo-10-hydroxycaminomycin, annamycin,galarubicin, elinafide, MEN10755 and4-demethoxy-3-deamino-3-aziridinyl-4-methylsulfonyldaunorubicin (see WO00/50032).

Examples of microtubulin inhibitors include paclitaxel, vindesinesulfate, 3′,4′-didehydro-4′-deoxy-8′-norvincaleukoblastine, docetaxol,rhizoxin, dolastatin, mivobulin isethionate, auristatin, cemadotin,RPR109881, BMS184476, vinflunine, cryptophycin,2,3,4,5,6-pentafluoro-N-(3-fluoro-4-methoxyphenyl)benzenesulfonamide,anhydrovinblastine,N,N-dimethyl-L-valyl-L-valyl-N-methyl-L-valyl-L-prolyl-L-proline-t-butylamide,TDX258 and BMS188797.

Some examples of topoisomerase inhibitors are topotecan, hycaptamine,irinotecan, rubitecan,6-ethoxypropionyl-3′,4′-O-exobenzylidene-chartreusin,9-methoxy-N,N-dimethyl-5-nitropyrazolo[3,4,5-kl]acridine-2-(6H)propanamine,1-amino-9-ethyl-5-fluoro-2,3-dihydro-9-hydroxy-4-methyl-1H,12H-benzo[de]pyrano[3′,4′:b,7]indolizino[1,2b]quinoline-10,13(9H,15H)dione,lurtotecan, 742-(N-isopropylamino)ethyl]-(20S)-camptothecin, BNP1350,BNPI1100, BN80915, BN80942, etoposide phosphate, teniposide, sobuzoxane,2′-dimethylamino-2′-deoxyetoposide, GL331,N42-(dimethylamino)ethyl]-9-hydroxy-5,6-dimethyl-6H-pyrido[4,3-b]carbazole-1-carboxamide,asulacrine,(5a,5aB,8aa,9b)-9-[2-[N-[2-(dimethylamino)ethyl]-methylamino]ethyl]-5-[4-hydroxy-3,5-dimethoxy-phenyl]-5,5a,6,8,8a,9-hexohydrofuro(3′,4′:6,7)naphtho(2,3-d)-1,3-dioxol-6-one,2,3-(methylenedioxy)-5-methyl-7-hydroxy-8-methoxybenzo[c]phen-anthridinium,6,9-bis[(2-aminoethyl)amino]benzo[g]isoquinoline-5,10-dione,5-(3-aminopropylamino)-7,10-dihydroxy-2-(2-hydroxyethylaminomethyl)-6H-pyrazolo[4,5,1-de]acridin-6-one,N-[1-[2(diethylamino)ethylamino]-7-methoxy-9-oxo-9H-thioxanthen-4-ylmethyl]formamide,N-(2-(dimethylamino)ethyl)acridine-4-carboxamide,6-[[2-(dimethylamino)ethyl]-amino]-3-hydroxy-7H-indeno[2,1-c]quinolin-7-oneand dimesna. “Antiproliferative agents” include antisense RNA and DNAoligonucleotides such as G3139, ODN698, RVASKRAS, GEM231 and INX3001 andantimetabolites such as enocitabine, carmofur, tegafur, pentostatin,doxifluridine, trimetrexate, fludarabine, capecitabine, galocitabine,cytarabine ocfosfate, fosteabine sodium hydrate, raltitrexed,paltitrexid, emitefur, tiazofurin, decitabine, nolatrexed, pemetrexed,nelzarabine, 2′-deoxy-2′-methylidenecytidine,2′-fluoromethylene-2′-deoxycytidine,N-[5-(2,3-dihydrobenzofuryl)sulfonyl]-N′-(3,4-dichlorophenyl)urea,N6-[4-deoxy-4-[N2-[2(E),4(E)-tetradecadienoyl]glycylamino]-L-glycero-B-L-manno-heptopyranosyl]adenine,aplidine, ecteinascidin, troxacitabine,4-[2-amino-4-oxo-4,6,7,8-tetrahydro-3H-pyrimidino[5,4-b][1,4]thiazin-6-yl-(S)-ethyl]-2,5-thienoyl-L-glutamicacid, aminopterin, 5-fluorouracil, alanosine,11-acetyl-8-(carbamoyloxymethyl)-4-formyl-6-methoxy-[4-oxa-1,1′-diaza-tetracyclo(7.4.1.0.0)tetradeca-2,4,6-trien-9-ylaceticacid ester, swainsonine, lometrexol, dexrazoxane, methioninase,2′-cyano-2′-deoxy-N4-palmitoyl-1-B-D-arabinofuranosyl cytosine and3-aminopyridine-2-carboxaldehyde thiosemicarbazone. “Antiproliferativeagents” also include monoclonal antibodies to growth factors other thanthose listed under “angiogenesis inhibitors”, such as trastuzumab andtumour suppressor genes, such as p53, which can be delivered viarecombinant virus-mediated gene transfer (see U.S. Pat. No. 6,069,134,for example).

As stated above the compounds according of the formula I are effectivemodulators (activators or inhibitors) of one or more protein kinasesselected from the group of Raf, Mek, PKB, Tie2, PDGFR and VEGFR.

Therefore, a preferred embodiment of the present invention are compoundsof formula I as protein kinase activators or inhibitors.

A particularly preferred embodiment of the present invention arecompounds of formula I as protein kinase activators or inhibitors,characterized in that the protein kinase is MEK1 or MEK2.

Thus, the compounds of the invention may also be useful as reagents forstudying signal transduction, protein kinases or any of the clinicaldisorders listed throughout this application.

For the identification of a signal transduction pathway and thedetection of cross talks with other signaling pathways suitable modelsor model systems have been generated by various scientists, for examplecell culture models (e.g. Khwaja et al., EMBO, 1997, 16, 2783-93) andtransgenic animal models (e.g. White et al., Oncogene, 2001, 20,7064-7072). For the examination of particular steps in the signaltransduction cascade, interfering compounds can be used for signalmodulation (e.g. Stephens et al., Biochemical J., 2000, 351, 95-105).The compounds according to the invention may also be useful as reagentsfor the examination of kinase dependent signal transduction pathways inanimal and/or cell culture models or any of the clinical disorderslisted throughout this application.

The measurement of kinase activity is a well-known technique feasiblefor each person skilled in the art. Generic test systems for kinaseactivity detection with substrates, for example histone (e.g. Alessi etal., FEBS Lett. 1996, 399, 3, page 333-8) or myelin basic protein arewell described in the literature (e.g. Campos-Gonzalez, R. and Glenney,Jr., J. R. 1992 J. Biol. Chem. 267, Page 14535).

For the identification of kinase inhibitors various assay systems areavailable (see for example Walters et al., Nature Drug Discovery 2003,2; page 259-266). For example, in scintillation proximity assays (e.g.Sorg et al., J. of. Biomolecular Screening, 2002, 7, 11-19) orflashplate assays the radioactive phosphorylation of a protein orpeptide as substrate with γATP can be measured. In the presence of aninhibitory compound no signal or a decreased radioactive signal isdetectable. Furthermore homogeneous time-resolved fluorescence resonanceenergy transfer (HTR-FRET), and fluorescence polarization (FP)technologies are useful for assay methods (for example Sills et al., J.of Biomolecular Screening, 2002: 191-214).

Other non-radioactive ELISA based assay methods use specificphospho-antibodies (AB). The phospho-AB binds only the phosphorylatedsubstrate. This binding is detectable with a secondary peroxidaseconjugated antibody, measured for example by chemiluminescence (forexample Ross et al., Biochem. J., 2002, 366: 977-981).

Other assays are known from the literature and could readily beperformed by the person skilled in the art (see, for example, Dhanabalet al., Cancer Res. 59:189-197; Xin et al., J. Biol. Chem.274:9116-9121; Sheu et al., Anticancer Res. 18:4435-4441; Ausprunk etal., Dev. Biol. 38:237-248; Gimbrone et al., J. Natl. Cancer Inst.52:413-427; Nicosia et al., In Vitro 18:538-549).

A further preferred embodiment of the present invention is apharmaceutical composition, characterized in that it contains atherapeutically effective amount of one or more compounds according tothe invention.

A further embodiment of the present invention is a pharmaceuticalcomposition, characterized in that it further contains one or moreadditional compounds, selected from the group consisting ofphysiologically acceptable excipients, auxiliaries, adjuvants, diluents,carriers and pharmaceutically active agents other than the compoundsaccording to the invention.

An additional preferred embodiment of the present invention is a set(kit) consisting of separate packets of

-   a) a therapeutically effective amount of one or more compounds    according to the invention and-   b) a therapeutically effective amount one or more further    pharmaceutically active agents other than the compounds according to    the invention.

A further embodiment of the present invention is a process for themanufacture of said pharmaceutical compositions, characterized in thatone or more compounds according to the invention and one or morecompounds selected from the group consisting of solid, liquid orsemiliquid excipients, auxiliaries, adjuvants, diluents, carriers andpharmaceutically active agents other than the compounds according to theinvention, are converted in a suitable dosage form.

The pharmaceutical compositions of the present invention may beadministered by any means that achieve their intended purpose. Forexample, administration may be by oral, parenteral, topical, enteral,intravenous, intramuscular, inhalant, nasal, intraarticular,intraspinal, transtracheal, transocular, subcutaneous, intraperitoneal,transdermal, or buccal routes. Alternatively, or concurrently,administration may be by the oral route. The dosage administered will bedependent upon the age, health, and weight of the recipient, kind ofconcurrent treatment, if any, frequency of treatment, and the nature ofthe effect desired. Parenteral administration is preferred. Oraladministration is especially preferred.

Suitable dosage forms include, but are not limited to capsules, tablets,pellets, dragees, semi-solids, powders, granules, suppositories,ointments, creams, lotions, inhalants, injections, cataplasms, gels,tapes, eye drops, solution, syrups, aerosols, suspension, emulsion,which can be produced according to methods known in the art, for exampleas described below:

Tablets:

mixing of active ingredients and auxiliaries, compression of saidmixture into tablets (direct compression), optionally granulation ofpart of mixture before compression.

Capsules:

mixing of active ingredient/s and auxiliaries to obtain a flowablepowder, optionally granulating powder, filling powders/granulate intoopened capsules, capping of capsules.

Semi-Solids (Ointments, Gels, Creams):

dissolving/dispersing active ingredient/s in an aqueous or fattycarrier; subsequent mixing of aqueous/fatty phase with complementaryfatty resp. aqueous phase, homogenization (creams only).

Suppositories (Rectal and Vaginal):

dissolving/dispersing active ingredient/s in carrier material liquifiedby heat (rectal: carrier material normally a wax; vaginal: carriernormally a heated solution of a gelling agent), casting said mixtureinto suppository forms, annealing and withdrawal suppositories from theforms.

Aerosols:

dispersing/dissolving active agents in a propellant, bottling saidmixture into an atomizer.

In general, non-chemical routes for the production of pharmaceuticalcompositions and/or pharmaceutical preparations comprise processingsteps on suitable mechanical means known in the art that transfer one ormore compounds according to the invention into a dosage form suitablefor administration to a patient in need of such a treatment. Usually,the transfer of one or more compounds according to the invention intosuch a dosage form comprises the addition of one or more compounds,selected from the group consisting of carriers, excipients, auxiliariesand pharmaceutical active ingredients other than the compounds accordingto the invention. Suitable processing steps include, but are not limitedto combining, milling, mixing, granulating, dissolving, dispersing,homogenizing, casting and/or compressing the respective active andnon-active ingredients. Mechanical means for performing said processingsteps are known in the art, for example from Ullmann's Encyclopedia ofIndustrial Chemistry, 5th Edition. In this respect, active ingredientsare preferably at least one compound according to this invention and oneor more additional compounds other than the compounds according to theinvention, which show valuable pharmaceutical properties, preferablythose pharmaceutical active agents other than the compounds according tothe invention, which are disclosed herein.

Particularly suitable for oral use are tablets, pills, coated tablets,capsules, powders, granules, syrups, juices or drops, suitable forrectal use are suppositories, suitable for parenteral use are solutions,preferably oil-based or aqueous solutions, furthermore suspensions,emulsions or implants, and suitable for topical use are ointments,creams or powders. The novel compounds may also be lyophilised and theresultant lyophilisates used, for example, for the preparation ofinjection preparations. The preparations indicated may be sterilisedand/or comprise assistants, such as lubricants, preservatives,stabilisers and/or wetting agents, emulsifiers, salts for modifying theosmotic pressure, buffer substances, dyes, flavours and/or a pluralityof further active ingredients, for example one or more vitamins.

Suitable excipients are organic or inorganic substances, which aresuitable for enteral (for example oral), parenteral or topicaladministration and do not react with the novel compounds, for examplewater, vegetable oils, benzyl alcohols, alkylene glycols, polyethyleneglycols, glycerol triacetate, gelatine, carbohydrates, such as lactose,sucrose, mannitol, sorbitol or starch (maize starch, wheat starch, ricestarch, potato starch), cellulose preparations and/or calciumphosphates, for example tricalcium phosphate or calcium hydrogenphosphate, magnesium stearate, talc, gelatine, tragacanth, methylcellulose, hydroxypropylmethylcellulose, sodium carboxymethylcellulose,polyvinyl pyrrolidone and/or Vaseline.

If desired, disintegrating agents may be added such as theabove-mentioned starches and also carboxymethyl-starch, cross-linkedpolyvinyl pyrrolidone, agar, or alginic acid or a salt thereof, such assodium alginate. Auxiliaries include, without limitation,flow-regulating agents and lubricants, for example, silica, talc,stearic acid or salts thereof, such as magnesium stearate or calciumstearate, and/or polyethylene glycol. Dragee cores are provided withsuitable coatings, which, if desired, are resistant to gastric juices.For this purpose, concentrated saccharide solutions may be used, whichmay optionally contain gum arabic, talc, polyvinyl pyrrolidone,polyethylene glycol and/or titanium dioxide, lacquer solutions andsuitable organic solvents or solvent mixtures. In order to producecoatings resistant to gastric juices or to provide a dosage formaffording the advantage of prolonged action, the tablet, dragee or pillcan comprise an inner dosage and an outer dosage component me latterbeing in the form of an envelope over the former. The two components canbe separated by an enteric layer, which serves to resist disintegrationin the stomach and permits the inner component to pass intact into theduodenum or to be delayed in release. A variety of materials can be usedfor such enteric layers or coatings, such materials including a numberof polymeric acids and mixtures of polymeric acids with such materialsas shellac, acetyl alcohol, solutions of suitable cellulose preparationssuch as acetyl-cellulose phthalate, cellulose acetate orhydroxypropymethyl-cellulose phthalate, are used. Dye stuffs or pigmentsmay be added to the tablets or dragee coatings, for example, foridentification or in order to characterize combinations of activecompound doses.

Suitable carrier substances are organic or inorganic substances whichare suitable for enteral (e.g. oral) or parenteral administration ortopical application and do not react with the novel compounds, forexample water, vegetable oils, benzyl alcohols, polyethylene glycols,gelatin, carbohydrates such as lactose or starch, magnesium stearate,talc and petroleum jelly. In particular, tablets, coated tablets,capsules, syrups, suspensions, drops or suppositories are used forenteral administration, solutions, preferably oily or aqueous solutions,furthermore suspensions, emulsions or implants, are used for parenteraladministration, and ointments, creams or powders are used for topicalapplication. The novel compounds can also be lyophilized and thelyophilizates obtained can be used, for example, for the production ofinjection preparations.

The preparations indicated can be sterilized and/or can containexcipients such as lubricants, preservatives, stabilizers and/or wettingagents, emulsifiers, salts for affecting the osmotic pressure, buffersubstances, colorants, flavourings and/or aromatizers. They can, ifdesired, also contain one or more further active compounds, e.g. one ormore vitamins.

Other pharmaceutical preparations, which can be used orally includepush-fit capsules made of gelatine, as well as soft, sealed capsulesmade of gelatine and a plasticizer such as glycerol or sorbitol. Thepush-fit capsules can contain the active compounds in the form ofgranules, which may be mixed with fillers such as lactose, binders suchas starches, and/or lubricants such as talc or magnesium stearate and,optionally, stabilizers. In soft capsules, the active compounds arepreferably dissolved or suspended in suitable liquids, such as fattyoils, or liquid paraffin. In addition, stabilizers may be added.

The liquid forms in which the novel compositions of the presentinvention may be incorporated for administration orally include aqueoussolutions, suitably flavoured syrups, aqueous or oil suspensions, andflavoured emulsions with edible oils such as cottonseed oil, sesame oil,coconut oil or peanut oil, as well as elixirs and similar pharmaceuticalvehicles. Suitable dispersing or suspending agents for aqueoussuspensions include synthetic and natural gums such as tragacanth,acacia, alginate, dextran, sodium carboxymethylcellulose,methylcellulose, polyvinyl-pyrrolidone or gelatine.

Suitable formulations for parenteral administration include aqueoussolutions of the active compounds in water-soluble form, for example,water-soluble salts and alkaline solutions. In addition, suspensions ofthe active compounds as appropriate oily injection suspensions may beadministered. Suitable lipophilic solvents or vehicles include fattyoils, for example, sesame oil, or synthetic fatty acid esters, forexample, ethyl oleate or triglycerides or polyethylene glycol-400 (thecompounds are soluble in PEG-400).

Aqueous injection suspensions may contain substances, which increase theviscosity of the suspension include, for example, sodium carboxymethylcellulose, sorbitol, and/or dextran, optionally, the suspension may alsocontain stabilizers.

For administration as an inhalation spray, it is possible to use spraysin which the active ingredient is either dissolved or suspended in apropellant gas or propellant gas mixture (for example CO₂ orchlorofluorocarbons). The active ingredient is advantageously used herein micronized form, in which case one or more additional physiologicallyacceptable solvents may be present, for example ethanol. Inhalationsolutions can be administered with the aid of conventional inhalers.

Possible pharmaceutical preparations, which can be used rectallyinclude, for example, suppositories, which consist of a combination ofone or more of the active compounds with a suppository base. Suitablesuppository bases are, for example, natural or synthetic triglycerides,or paraffin hydrocarbons. In addition, it is also possible to usegelatine rectal capsules, which consist of a combination of the activecompounds with a base. Possible base materials include, for example,liquid triglycerides, polyethylene glycols, or paraffin hydrocarbons.

For use in medicine, the compounds of the present invention will be inthe form of pharmaceutically acceptable salts. Other salts may, however,be useful in the preparation of the compounds according to the inventionor of their pharmaceutically acceptable salts. Suitable pharmaceuticallyacceptable salts of the compounds of this invention include acidaddition salts which may, for example be formed by mixing a solution ofthe compound according to the invention with a solution of apharmaceutically acceptable acid such as hydrochloric acid, sulphuricacid, methanesulphonic acid, fumaric acid, maleic acid, succinic acid,acetic acid, benzoic acid, oxalic acid, citric acid, tartaric acid,carbonic acid or phosphoric acid. Furthermore, where the compounds ofthe invention carry an acidic moiety, suitable pharmaceuticallyacceptable salts thereof may include alkali metal salts, e.g. sodium orpotassium salts; alkaline earth metal salts, e.g. calcium or magnesiumsalts; and salts formed with suitable organic bases, e.g. quaternaryammonium salts.

The present invention includes within its scope prodrugs of thecompounds of the present invention above. In general, such prodrugs willbe functional derivatives of the compounds of the present invention,which are readily convertible in vivo into the required compound of thepresent invention. Conventional procedures for the selection andpreparation of suitable prodrug derivatives are described, for example,in Design of Prodrugs, ed. H. Bundgaard, Elsevier, 1985.

The pharmaceutical preparations can be employed as medicaments in humanand veterinary medicine. As used herein, the term “effective amount”means that amount of a drug or pharmaceutical agent that will elicit thebiological or medical response of a tissue, system, animal or human thatis being sought, for instance, by a researcher or clinician.Furthermore, the term “therapeutically effective amount” means anyamount which, as compared to a corresponding subject who has notreceived such amount, results in improved treatment, healing,prevention, or amelioration of a disease, disorder, or side effect, or adecrease in the rate of advancement of a disease or disorder. The termalso includes within its scope amounts effective to enhance normalphysiological function. Said therapeutic effective amount of one or moreof the compounds according to the invention is known to the skilledartisan or can be easily determined by standard methods known in theart.

The substances according to the invention are generally administeredanalogously to commercial preparations. Usually, suitable doses that aretherapeutically effective lie in the range between 0.0005 mg and 1000mg, preferably between 0.005 mg and 500 mg and especially between 0.5and 100 mg per dose unit. The daily dose is preferably between about0.001 and 10 mg/kg of body weight.

Those of skill will readily appreciate that dose levels can vary as afunction of the specific compound, the severity of the symptoms and thesusceptibility of the subject to side effects. Some of the specificcompounds are more potent than others. Preferred dosages for a givencompound are readily determinable by those of skill in the art by avariety of means. A preferred means is to measure the physiologicalpotency of a given compound.

The host, or patient, may be from any mammalian species, e.g., primatesp., particularly human; rodents, including mice, rats and hamsters;rabbits; equines, bovines, canines, felines; etc. Animal models are ofinterest for experimental investigations, providing a model fortreatment of human disease.

The specific dose for the individual patient depends, however, on themultitude of factors, for example on the efficacy of the specificcompounds employed, on the age, body weight, general state of health,the sex, the kind of diet, on the time and route of administration, onthe excretion rate, the kind of administration and the dosage form to beadministered, the pharmaceutical combination and severity of theparticular disorder to which the therapy relates. The specifictherapeutic effective dose for the individual patient can readily bedetermined by routine experimentation, for example by the doctor orphysician, which advises or attends the therapeutic treatment.

In the case of hyperproliferative disorders, the susceptibility of aparticular cell to treatment with the subject compounds may bedetermined by in vitro testing. Typically a culture of the cell iscombined with a subject compound at varying concentrations for a periodof time sufficient to allow the active agents to induce cell death orinhibit migration, usually between about one hour and one week. For invitro testing, cultured cells from a biopsy sample may be used. Theviable cells, left after treatment, are then counted.

The dose will vary depending on the specific compound utilized, specificdisorder, patient status, etc. Typically a therapeutic dose will besufficient to substantially decrease the undesirable cell population inthe targeted tissue, while maintaining patient viability. Treatment willgenerally be continued until there is a substantial reduction, e.g., atleast about 50%, decrease in the cell burden, and may be continued untilthere are essentially none of the undesirable cells detected in thebody.

Even without further details, it is assumed that a person skilled in theart will be able to utilise the above description in the broadest scope.The preferred embodiments should therefore merely be regarded asdescriptive disclosure, which is absolutely not limiting in any way.

Above and below, all temperatures are indicated in ° C. In the followingexamples, “conventional work-up” means that, if necessary, the solventis removed, water is added if necessary, the pH is adjusted, ifnecessary, to between 2 and 10, depending on the constitution of the endproduct, the mixture is extracted with ethyl acetate or dichloromethane,the phases are separated, the organic phase is washed with saturatedNaHCO₃ solution, if desired with water and saturated NaCl solution, isdried over sodium sulfate, filtered and evaporated, and the product ispurified by chromatography on silica gel, by preparative HPLC and/or bycrystallisation. The purified compounds are, if desired, freeze-dried.

Mass spectrometry (MS): ESI (electrospray ionisation) (M+H)⁺

List of Abbreviations and Acronyms:

AcOH acetic acid, anh anhydrous, atm atmosphere(s), BOCtert-butoxycarbonyl CDI 1,1′-carbonyl diimidazole, cone concentrated, dday(s), dec decomposition, DMAC N,N-dimethylacetamide, DMPU1,3-dimethyl-3,4,5,6-tetrahydro-2(IH)-pyrimidinone, DMFN,N-dimethylformamide, DMSO dimethylsulfoxide, DPPA diphenylphosphorylazide, EDCl 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide, EtOAc ethylacetate, EtOH ethanol (100%), Et₂Q diethyl ether, Et₃N triethylamine, hhour(s), MeOH methanol, pet. ether petroleum ether (boiling range 30-60°C.), temp. temperature, THF tetrahydrofuran, TFA trifluoroAcOH, Tftrifluoromethanesulfonyl.

EXAMPLE 1 Preparation of Amino-Oxindoles

One equivalent of a substituted aromatic or heteroaromatic nitrile (herea compound of formula II, wherein R¹ is phenyl, R⁷ is I, X is (CH₂)_(p)and p=0) is dissolved in 1,4-dioxane. After addition of 1,3 equivalentsof a straight or branched chain alcohol of formula III (wherein R⁹ is(CH₂)_(q) and q is 1-10), HCl-gas is introduced at 10° C. for 6 h. Theresulting pellet (compound of formula IV) is filtered, washed withdioxane and dried in vacuum.

IVa) 3-Iodo-benzimidic acid ethyl ester hydrochloride: 6.05 g (81%),colourless powder; HPLC: 2.11 min; LC-MS: 262.0 m/z.

IVb) 4-Iodo-benzimidic acid ethyl ester hydrochloride: 4.80 g (62%),beige crystals; HPLC: 2.00 min; LC-MS: 262.0 m/z.

Triethylamine (3 eq.), the compound of formula IV (2 eq.) and thecompound of formula V (1 eq.) are dissolved in 1-butanol (7 eq.) and areirradiated for 70 min at 200° C. in a microwave. The resulting product(respective compound of formula I, see following table) is purified bycolumn chromatography.

Retention time Amount/ Yield/ HPLC/ LC-MS/ No. Structure mg % min mz-1IC₅₀ a

37 22 2.77 271.0 1.10E-06 b

47 29 2.75 271.0 2.60E-07 c

14 7 2.33 253.0 9.40E-07 d

16 6 2.83 363.0 2.70E-06 e

37 15 3.00 397.0 4.10E-06 f

19 8 2.95 397.0 1.10E-06 g

44 16 2.79 363.0 n.d. h

30 13 2.95 397.0 n.d. i

44 19 2.91 397.0 n.d. j

43 17 2.97 377.0 n.d. k

33 22 2.77 316.0 6.60E-07 l

25 10 2.95 377.0 n.d. m

24 11 2.95 440.9 n.d. n

20 10 2.95 440.8 3.40E-06 o

21 10 2.59 267.2 3.10E-06 p

37 20 2.72 301.0 8.70E-06 q

25 15 2.73 345.0 n.d. r

98 19 2.49 287.0 n.d. s

260 51 2.47 287.0 n.d. t

59 12 2.71 251.2 n.d. u

70 13 2.48 267.2 n.d. v

64 14 2.52 331.0 n.d. w

97 18 2.67 301.0 n.d. x

47 12 2.6 255.2 n.d. y

51 15 2.73 289.0 n.d. z

31 8 2.67 281.2 n.d.

EXAMPLE 2 Cellular Assay for Measuring MEK Inhibition

MEK inhibitors are evaluated by determining their ability to inhibitphosphorylation of MAP kinase (ERK) in murine colon 26 (C26) carcinomacells. Since ERK1 and ERK2 represent the only known substrates for MEK1and MEK2, the measurement of inhibition of ERK phosphorylation in cellsprovides direct read out of cellular MEK inhibition by the compounds ofthe invention. Detection of phosphorylation of ERK is carried out eitherby Western blot or ELISA format. Briefly, the assays involve treatmentof exponentially growing C26 cells with varying concentrations of thetest compound (or vehicle control) for one hour at 37° C. For Westernblot assay, cells are rinsed free of compound/vehicle and lysed in asolution containing 70 mM NaCl, 50 mM glycerol phosphate, 10 mM HEPES,pH 7.4, 1% Triton X-100, 1 mM Na₃VO₄, 100 μM PMSF, 10 μM leupeptin and10 μM pepstatin. Supernatants were then subjected to gel electrophoresisand hybridized to a primary antibody recognizing dually phosphorylatedERKl and ERK2. To evaluate total MAPK levels, blots were subsequently‘stripped’ and re-probed with a 1:1 mixture of polyclonal antibodiesrecognizing unphosphorylated ERK1 and ERK2. For pERK ELISA assay, pERKTiterZyme Enzyme immunometric Assay kits were acquired from AssayDesigns, Inc (Ann Arbor, Mich.). Briefly, cells were harvested in lysissolution containing 50 mM B-glycerophosphate, 10 mM HEPES, pH 7.4, 70 mMNaCl, 2 mM EDTA and 1% SDS and protein lysates were diluted 1:15 withsupplied Assay buffer prior to the execution of the assay. Thesubsequent steps were carried out essentially as recommended by themanufacturer.

EXAMPLE 3 VEGF Receptor Kinase Assay

VEGF receptor kinase activity is measured by incorporation ofradio-labelled phosphate into polyglutamic acid/tyrosine, 4:1 (pEY)substrate. The phosphorylated pEY product is trapped onto a filtermembrane and the incorporation of radiolabelled phosphate is quantifiedby scintillation counting.

Materials

VEGF receptor kinase: The intracellular tyrosine kinase domains of humanKDR (Terman, B. I. et al. Oncogene (1991) Vol. 6, pp. 1677-1683.) andFlt-1 (Shibuya, M. et al. Oncogene (1990) Vol. 5, pp. 519-524) werecloned as glutathione S-transferase (GST) gene fusion proteins. This wasaccomplished by cloning the cytoplasmic domain of the KDR kinase as anin frame fusion at the carboxyl terminus of the GST gene. Solublerecombinant GST-kinase domain fusion proteins are expressed inSpodoptera frugiperda (Sf21) insect cells (Invitrogen) using abaculovirus expression vector (pAcG2T, Pharmingen).

Lysis buffer: 50 mM Tris pH 7.4, 0.5 M NaCl, 5 mM DTT, 1 mM EDTA, 0.5%triton X-100, 10% glycerol, 10 mg/ml of each leupeptin, pepstatin andaprotinin and 1 mM phenylmethylsulfonyl fluoride (all Sigma).

Wash buffer: 50 mM Tris pH 7.4, 0.5 M NaCl, 5 mM DTT, 1 mM EDTA, 0.05%triton X-100, 10% glycerol, 10 mg/ml of each leupeptin, pepstatin andaprotinin and 1 mM phenylmethylsulfonyl fluoride.

Dialysis buffer: 50 mM Tris pH 7.4, 0.5 M NaCl, 5 mM DTT, 1 mM EDTA,0.05% triton X-100, 50% glycerol, 10 mg/ml of each leupeptin, pepstatinand aprotinin and 1 mM phenylmethylsulfonyl fluoride.

10× reaction buffer: 200 mM Tris, pH 7.4, 1.0 M NaCl, 50 mM MnCl₂, 10 mMDTT and 5 mg/ml bovine serum albumin [BSA] (Sigma).

Enzyme dilution buffer: 50 mM Tris, pH 7.4, 0.1 M NaCl, 1 mM DTT, 10%glycerol, 100 mg/ml BSA.

10× substrate: 750 μg/ml poly(glutamic acid/tyrosine; 4:1) (Sigma).

Stop solution: 30% trichloroacetic acid, 0.2 M sodium pyrophosphate(both Fisher).

Wash solution: 15% trichloroacetic acid, 0.2 M sodium pyrophosphate.

Filter Plates

Millipore #MAFC NOB, GF/C glass fibre 96 well plate.

Method A—Protein Purification

1. Sf21 cells are infected with recombinant virus at a multiplicity ofinfection of 5 virus particles/cell and grown at 27° C. for 48 hours.

2. All steps are performed at 4° C. Infected cells are harvested bycentrifugation at 1000×g and lysed at 4° C. for 30 minutes with 1/10volume of lysis buffer followed by centrifugation at 100,000×g for 1hour. The supernatant is then passed over a glutathione Sepharose column(Pharmacia) equilibrated with lysis buffer and washed with 5 volumes ofthe same buffer followed by 5 volumes of wash buffer. RecombinantGST-KDR protein is eluted with wash buffer/10 mM reduced glutathione(Sigma) and dialysed against dialysis buffer.

Method B—VEGF Receptor Kinase Assay

1, Add 5 μl of inhibitor or control to the assay in 50% DMSO.

2. Add 35 μl of reaction mixture containing 5 μl of 10× reaction buffer,5 μl 25 mM ATP/10 μCi [³³P]ATP (Amersham) and 5 μl of 10× substrate.

3. Start the reaction by the addition of 10 μl of KDR (25 nM) in enzymedilution buffer.

4. Mix and incubate at room temperature for 15 minutes.

5. Stop the reaction by the addition of 50 μl of stop solution.

6. Incubate for 15 minutes at 4° C.

7. Transfer a 90 μl aliquot to filter plate.

8. Aspirate and wash 3 times with wash solution.

9. Add 30 μl of scintillation cocktail, seal plate and count in aWallace Microbeta scintillation counter.

Human Umbilical Vein Endothelial Cell Mitogenesis Assay

Expression of VEGF receptors that mediate mitogenic responses to thegrowth factor is largely restricted to vascular endothelial cells. Humanumbilical vein endothelial cells (HUVECs) in culture proliferate inresponse to VEGF treatment and can be used as an assay system toquantify the effects of KDR kinase inhibitors on VEGF stimulation. Inthe assay described, quiescent HUVEC monolayers are treated with vehicleor test compound 2 hours prior to addition of VEGF or basic fibroblastgrowth factor (BFGF). The mitogenic response to VEGF or BFGF isdetermined by measuring the incorporation of [³H]thymidine into cellularDNA.

Materials

HUVECs: HUVECs frozen as primary culture isolates are obtained fromClonetics Corp. Cells are maintained in endothelial growth medium (EGM;Clonetics) and are used for mitogenic assays at passages 3-7.

Culture plates: NUNCLON 96-well polystyrene tissue culture plates (NUNC#167008).

Assay medium: Dulbecco's modification of Eagle's medium containing 1g/ml glucose (low-glucose DMEM; Mediatech) plus 10% (v/v) foetal bovineserum (Clonetics).

Test compounds: Working stock solutions of test compounds are dilutedserially in 100% dimethyl sulfoxide (DMSO) to 400 times greater thantheir desired final concentrations. Final dilutions to 1× concentrationare made directly into assay medium immediately prior to addition tocells.

10× growth factors: Solutions of human VEGF 165 (500 ng/ml; R&D Systems)and BFGF (10 ng/ml; R&D Systems) are prepared in assay medium.

10×[³H]thymidine: [Methyl-3H]thymidine (20 Ci/mmol; Dupont-NEN) isdiluted to 80 μCi/ml in low-glucose DMEM.

Cell wash medium: Hank's balanced salt solution (Mediatech) containing 1mg/ml bovine serum albumin (Boehringer-Mannheim).

Cell lysis solution: 1 N NaOH, 2% (w/v) Na₂CO₃.

Method 1

HUVEC monolayers maintained in EGM are harvested by trypsinisation andplated out at a density of 4000 cells per 100 μl of assay medium perwell in 96-well plates. Cells growth is arrested for 24 hours at 37° C.in a humidified atmosphere containing 5% CO₂.

Method 2

Growth-arrest medium is replaced by 100 μl of assay medium containingeither vehicle (0.25% [v/v] DMSO) or the desired final concentration oftest compound. All determinations are performed in triplicate. Cells arethen incubated at 37° C./5% CO₂ for 2 hours to allow test compounds toenter cells.

Method 3

After the 2-hour pre-treatment period, cells are stimulated by additionof 10 μl/well of either assay medium, 10×VEGF solution or 10×BFGFsolution. Cells are then incubated at 37° C./5% CO₂.

Method 4

After 24 hours in the presence of growth factors, 10×[³H]thymidine (10μl/well) is added.

Method 5

Three days after addition of [³H]thymidine, medium is removed byaspiration, and cells are washed twice with cell wash medium (400μl/well followed by 200 μl/well). The washed, adherent cells are thensolubilised by addition of cell lysis solution (100 μl/well) and warmingto 37° C. for 30 minutes. Cell lysates are transferred to 7 ml glassscintillation vials containing 150 μl of water. Scintillation cocktail(5 ml/vial) is added, and cell-associated radioactivity is determined byliquid scintillation spectroscopy. According to these assays, thecompounds of the formula I are inhibitors of VEGF and are thus suitablefor the inhibition of angiogenesis, such as in the treatment of oculardiseases, for example diabetic retinopathy, and for the treatment ofcarcinomas, for example solid tumours. The present compounds inhibitVEGF-stimulated mitogenesis of human vascular endothelial cells inculture with IC 50 values of 0.01-5.0 μM. These compounds also showselectivity over related tyrosine kinases (for example, FGFR1 and theSrc family; for relationship between Src kinases and VEGFR kinases, seeEliceiri et al., Molecular Cell, Vol. 4, pp. 915-924, December 1999).

EXAMPLE 4 Injection Vials

A solution of 100 g of an active compound of the present invention and 5g of disodium hydrogenphosphate is adjusted to pH 6.5 in 3 l ofdouble-distilled water using 2N hydrochloric acid, sterile-filtered,dispensed into injection vials, lyophilized under sterile conditions andaseptically sealed. Each injection vial contains 5 mg of activecompound.

EXAMPLE 5 Suppositories

A mixture of 20 g of an active compound of the present invention isfused with 100 g of soya lecithin and 1400 g of cocoa butter, pouredinto moulds and allowed to cool. Each suppository contains 20 mg ofactive compound.

EXAMPLE 6 Solution

A solution of 1 g of an active compound of the present invention, 9.38 gof NaH₂PO₄.2H₂O, 28.48 g of Na₂HPO₄.12H₂O and 0.1 g of benzalkoniumchloride in 940 ml of double-distilled water is prepared. It is adjustedto pH 6.8, made up to 1 L and sterilized by irradiation. This solutioncan be used in the form of eye drops.

EXAMPLE 7 Ointment

500 mg of an active compound of the present invention is mixed with 99.5g of petroleum jelly under aseptic conditions.

EXAMPLE 8 Tablets

A mixture of 1 kg of active compound of the present invention, 4 kg oflactose, 1.2 kg of potato starch, 0.2 kg of talc and 0.1 kg of magnesiumstearate is compressed to give tablets in a customary manner such thateach tablet contains 10 mg of active compound.

EXAMPLE 9 Coated Tablets

Analogously to Example E, tablets are pressed and are then coated in acustomary manner using a coating of sucrose, potato starch, talc,tragacanth and colourant.

EXAMPLE 10 Capsules

2 kg of active compound of the present invention are dispensed into hardgelatin capsules in a customary manner such that each capsule contains20 mg of the active compound.

1-28. (canceled)
 29. A method for activating or inhibiting proteinkinase is MEK1 or MEK2, or for treating or preventing cancer, or fortreating or preventing melanoma, brain cancer, lung cancer, non-smallcell lung carcinoma, squamous cell cancer, colon cancer, duodenalcancer, ductal cancer, colorectal cancer, gastric cancer, stomachcancer, pancreatic cancer, hepatic cancer, renal cancer, bladder cancer,endometrial cancer, ovarian cancer, uterine cancer, prostate cancer,breast cancer, head cancer, neck cancer, oesophageal cancer,gynecological cancer, dysplastic oral mucosa, polyposis, invasive oralcancer, thyroid cancer, lymphoma, chronic leukaemia or acute leukaemia,comprising administering to a subject in need thereof an effectiveamount of a compound of formula I

wherein X is (CH₂)_(p). R¹ is Ar or Het, R² is H, A, Ar,(CH₂)_(m)CON(R⁸)₂ or (CH₂)_(m)CONHAr, R³, R⁴, R⁶, and R⁷ areindependently from each other H, A, Ar, OR⁸, SR^(a), OAr, SAr, N(R⁸)₂,NHAr, NAr₂, Hal, NO₂, CN, COR^(S), COAr, NHCOA, NHCOAr, NHSO₂A, NHSO₂Ar,SO₂N(R⁸)₂, O(CH₂)_(n)N(R⁸)₂ or O(CH₂)_(n)NHR⁸, R⁵ is H, A, Ar, OR⁸, SR⁸,OAr, SAr, N(R⁸)₂, NHAr, NAr₂, Hal, NO₂, CN, COAr, NHCOA, NHCOAr, NHSO₂A,NHSO₂Ar, SO₂N(R⁸)₂, O(CH₂),N(R⁵)₂ or O(CH₂)_(n)NHR⁸, R⁸ is H, A or A-Ar,A is a linear or branched alkyl or a cycloalkyl which is optionallysubstituted by Hal, Ar is aryl, Het is heteroaryl, Hal Cl, Br, I or F,n, and p are independently from each other 0-5, and m is 0-2, with theprovisio, that one of the residues R², R³, R⁴, R⁵, R⁶ or R⁷ is otherthan H and that3-(1-Amino-2-phenyl-ethylidene)-1-methyl-1,3-dihydro-indol-2-one isexcluded, or a pharmaceutically acceptable salt, derivative, prodrug,solvate or stereoisomer thereof, or a mixtures thereof.
 30. A methodaccording to claim 29, wherein in the compound of formula I X is(CH₂)_(p), p is 0-5, R¹ is Ar or Het, R², R³, R⁴, R⁶, and R⁷ are H, andR⁵ is Hal.
 31. A method according to claim 29, wherein in the compoundof formula I X is (CH₂)_(p), p is 0-5, R¹ is Ar or Het, R², R³, R⁵, R⁶,and R⁷ are H, and R⁴ is Hal.
 32. A method according to claim 29, whereinin the compound of formula I X is (CH₂)_(p), p is 0-5, R¹ is Ar or Het,R³, R⁴, R⁵, R⁶, and R⁷ are H, and R² is A or Ar.
 33. A method accordingto claim 29, wherein in the compound of formula I X is (CH₂)_(p), p is0-5, R¹ is Ar or Het, R², R³, R⁴, and R⁶ are H, R⁵ is Hal, R⁷ is Hal orOR⁸, and R⁸ is H or A.
 34. A method according to claim 29, wherein inthe compound of formula I X is (CH₂)_(p), p is 0-5, R¹ is Ar or Het, R²,R³, R⁵, and R⁶ are H, R⁴ is Hal, R⁷ is Hal or OR⁸, and R⁸ is H or A. 35.A method according to claim 29, wherein in the compound of formula I Xis (CHO₂)_(p), p is 0-5, R¹ is Ar or Het, R², R³, R⁴, and R⁶ are H, R⁵is Hal, R⁷ is Hal or OR⁸, and R⁸ is H or A.
 36. A method according toclaim 29, wherein in the compound of formula I X is (CH₂)_(p), p is 0,R¹ is phenyl, R³, R⁴, R⁵, and R⁶ are H, R² is A or Ar, R⁷ is Hal or OR⁸,and R⁸ is H or A.
 37. A method according to claim 29, wherein a compoundof formula I or a pharmaceutically acceptable salt thereof isadministered.
 38. A method according to claim 29, which is foractivating or inhibiting protein kinase is MEK1 or MEK2.
 39. A methodaccording to claim 29, which is for treating or preventing cancer.
 40. Amethod according to claim 29, which is for treating or preventingmelanoma, brain cancer, lung cancer, non-small cell lung carcinoma,squamous cell cancer, colon cancer, duodenal cancer, ductal cancer,colorectal cancer, gastric cancer, stomach cancer, pancreatic cancer,hepatic cancer, renal cancer, bladder cancer, endometrial cancer,ovarian cancer, uterine cancer, prostate cancer, breast cancer, headcancer, neck cancer, oesophageal cancer, gynecological cancer,dysplastic oral mucosa, polyposis, invasive oral cancer, thyroid cancer,lymphoma, chronic leukaemia or acute leukaemia.
 41. A method accordingto claim 29, which is for treating melanoma, brain cancer, lung cancer,non-small cell lung carcinoma, squamous cell cancer, colon cancer,duodenal cancer, ductal cancer, colorectal cancer, gastric cancer,stomach cancer, pancreatic cancer, hepatic cancer, renal cancer, bladdercancer, endometrial cancer, ovarian cancer, uterine cancer, prostatecancer, breast cancer, head cancer, neck cancer, oesophageal cancer,gynecological cancer, dysplastic oral mucosa, polyposis, invasive oralcancer, thyroid cancer, lymphoma, chronic leukaemia or acute leukaemia.42. A method according to claim 29, which further comprisesadministering an estrogen receptor modulator, androgen receptormodulator, retinoid receptor modulator, cytotoxic agent,anti-proliferative agent, prenyl protein protease inhibitor, HMG CoAreductase inhibitor, HIV protease inhibitor, reverse transcriptaseinhibitor, growth factor receptor inhibitor or angiogenesis inhibitor.43. A method according to claim 29, which further comprisesadministering an anti-metastatic, antitumor or anti-angiogenic agent,which is not a compound of formula I.
 44. A method according to claim29, which further comprises providing radio therapy to the subject inneed thereof.
 45. A method for activating or inhibiting protein kinaseis MEK1 or MEK2, or for treating or preventing cancer, or for treatingor preventing melanoma, brain cancer, lung cancer, non-small cell lungcarcinoma, squamous cell cancer, colon cancer, duodenal cancer, ductalcancer, colorectal cancer, gastric cancer, stomach cancer, pancreaticcancer, hepatic cancer, renal cancer, bladder cancer, endometrialcancer, ovarian cancer, uterine cancer, prostate cancer, breast cancer,head cancer, neck cancer, oesophageal cancer, gynecological cancer,dysplastic oral mucosa, polyposis, invasive oral cancer, thyroid cancer,lymphoma, chronic leukaemia or acute leukaemia, comprising administeringto a subject in need thereof an effective amount of a compound selectedfrom the group consisting of a)3-(Amino-phenyl-methylene)-6-chloro-1,3-dihydro-indol-2-one, b)3-(Amino-phenyl-methylene)-5-chloro-1,3-dihydro-indol-2-one, c)3-[Amino-(4-hydroxy-phenyl)methylene]-1,3-dihydro-indol-2-one, d)3-[Amino-(4-iodo-phenyl)methylene]-1,3-dihydro-indol-2-one, e)3-(Amino-(4-iodo-phenyl)-methylene)-6-chloro-1,3-dihydro-indol-2-one, f)3-(Amino-(4-iodo-phenyl)-methylene)-5-chloro-1,3-dihydro-indol-2-one, g)3-[Amino-(3-iodo-phenyl)-methylene]-1,3-dihydro-indol-2-one, h)3-(Amino-(3-iodo-phenyl)-methylene)-6-chloro-1,3-dihydro-indol-2-one, i)3-(Amino-(3-iodo-phenyl)-methylene)-5-chloro-1,3-dihydro-indol-2-one, j)3-(Amino-(3-iodo-phenyl)-methylene)-1-methyl-1,3-dihydro-indol-2-one, k)3-(Amino-phenyl-methylene)-5-bromo-1,3-dihydro-indol-2-one, l)3-(Amino-(4-iodo-phenyl)-methylene)-1-methyl-1,3-dihydro-indol-2-one, m)3-(Amino-(4-iodo-phenyl)-methylene)-5-bromo-1,3-dihydro-indol-2-one, n)3-(Amino-(3-iodo-phenyl)-methylene)-5-bromo-1,3-dihydro-indol-2-one, o)3-[Amino-(4-methoxy-phenyl)-methylene]-1,3-dihydro-indol-2-one, p)3-(Amino-(4-methoxy-phenyl)-methylene)-6-chloro-1,3-dihydro-indol-2-one,q)3-(Amino-(4-methoxy-phenyl)-methylene)-5-bromo-1,3-dihydro-indol-2-one,r)3-(Amino-(4-hydroxy-phenyl)-methylene)-6-chloro-1,3-dihydro-indol-2-one,s)3-(Amino-(4-hydroxy-phenyl)-methylene)-5-chloro-1,3-dihydro-indol-2-one,t) 3-(Amino-phenyl-methylene)-1-methyl-1,3-dihydro-indol-2-one, u)3-(Amino-(4-hydroxy-phenyl)-methylene)-1-methyl-1,3-dihydro-indol-2-one,v)3-(Amino-(4-hydroxy-phenyl)-methylene)-5-bromo-1,3-dihydro-indol-2-one,w)3-(Amino-(4-methoxy-phenyl)-methylene)-5-chloro-1,3-dihydro-indol-2-one,x) 3-[Amino-(4-fluoro-phenyl)methylene]-1,3-dihydro-indol-2-one, y)3-(Amino-(4-fluoro-phenyl)-methylene)-5-chloro-1,3-dihydro-indol-2-one,and z)3-(Amino-(4-methoxy-phenyl)-methylene)-1-methyl-1,3-dihydro-indol-2-one,and the physiologically acceptable salts, derivatives, prodrugs,solvates and stereoisomers thereof, including mixtures thereof in allratios.
 46. A method according to claim 45, wherein a)3-(Amino-phenyl-methylene)-6-chloro-1,3-dihydro-indol-2-one, b)3-(Amino-phenyl-methylene)-5-chloro-1,3-dihydro-indol-2-one, c)3-[Amino-(4-hydroxy-phenyl)-methylene]-1,3-dihydro-indol-2-one, d)3-[Amino-(4-iodo-phenyl)-methylene]-1,3-dihydro-indol-2-one, e)3-(Amino-(4-iodo-phenyl)-methylene)-6-chloro-1,3-dihydro-indol-2-one, f)3-(Amino-(4-iodo-phenyl)-methylene)-5-chloro-1,3-dihydro-indol-2-one, g)3-[Amino-(3-iodo-phenyl)-methylene]-1,3-dihydro-indol-2-one, h)3-(Amino-(3-iodo-phenyl)-methylene)-6-chloro-1,3-dihydro-indol-2-one, i)3-(Amino-(3-iodo-phenyl)-methylene)-5-chloro-1,3-dihydro-indol-2-one, j)3-(Amino-(3-iodo-phenyl)-methylene)-1-methyl-1,3-dihydro-indol-2-one, k)3-(Amino-phenyl-methylene)-5-bromo-1,3-dihydro-indol-2-one, l)3-(Amino-(4-iodo-phenyl)-methylene)-1-methyl-1,3-dihydro-indol-2-one, m)3-(Amino-(4-iodo-phenyl)-methylene)-5-bromo-1,3-dihydro-indol-2-one, n)3-(Amino-(3-iodo-phenyl)-methylene)-5-bromo-1,3-dihydro-indol-2-one, o)3-(Amino-(4-methoxy-phenyl)-methylene]-1,3-dihydro-indol-2-one, p)3-(Amino-(4-methoxy-phenyl)-methylene)-6-chloro-1,3-dihydro-indol-2-one,q)3-(Amino-(4-methoxy-phenyl)methylene)-5-bromo-1,3-dihydro-indol-2-one,r)3-(Amino-(4-hydroxy-phenyl)-methylene)-6-chloro-1,3-dihydro-indol-2-one,s)3-(Amino-(4-hydroxy-phenyl)methylene)-5-chloro-1,3-dihydro-indol-2-one,t) 3-(Amino-phenyl-methylene)-1-methyl-1,3-dihydro-indol-2-one, u)3-(Amino-(4-hydroxy-phenyl)-methylene)-1-methyl-1,3-dihydro-indol-2-one,v)3-(Amino-(4-hydroxy-phenyl)-methylene)-5-bromo-1,3-dihydro-indol-2-one,w)3-(Amino-(4-methoxy-phenyl)-methylene)-5-chloro-1,3-dihydro-indol-2-one,x) 3-(Amino-(4-fluoro-phenyl)-methylene]-1,3-dihydro-indol-2-one, y)3-(Amino-(4-fluoro-phenyl)-methylene)-5-chloro-1,3-dihydro-indol-2-one,or z)3-(Amino-(4-methoxy-phenyl)-methylene)-1-methyl-1,3-dihydro-indol-2-one,or a physiologically acceptable salt thereof is administered.
 47. Amethod according to claim 45, which is for treating melanoma, braincancer, lung cancer, non-small cell lung carcinoma, squamous cellcancer, colon cancer, duodenal cancer, ductal cancer, colorectal cancer,gastric cancer, stomach cancer, pancreatic cancer, hepatic cancer, renalcancer, bladder cancer, endometrial cancer, ovarian cancer, uterinecancer, prostate cancer, breast cancer, head cancer, neck cancer,oesophageal cancer, gynecological cancer, dysplastic oral mucosa,polyposis, invasive oral cancer, thyroid cancer, lymphoma, chronicleukaemia or acute leukaemia.